Human kinases

Abstract

The invention provides human kinases (PKIN) and polynucleotides which identify and encode PKIN. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating, or preventing disorders associated with aberrant expression of PKIN.

Description

TECHNICAL FIELD

[0001] This invention relates to nucleic acid and amino acid sequences of human kinases and to the use of these sequences in the diagnosis, treatment, and prevention of cancer, immune disorders, disorders affecting growth and development, cardiovascular diseases, and lipid disorders, and in the assessment of the effects of exogenous compounds on the expression of nucleic acid and amino acid sequences of human kinases.

BACKGROUND OF THE INVENTION

[0002] Kinases comprise the largest known enzyme superfamily and vary widely in their target molecules. Kinases catalyze the transfer of high energy phosphate groups from a phosphate donor to a phosphate acceptor. Nucleotides usually serve as the phosphate donor in these reactions, with most kinases utilizing adenosine triphosphate (ATP). The phosphate acceptor can be any of a variety of molecules, including nucleosides, nucleotides, lipids, carbohydrates, and proteins. Proteins are phosphorylated on hydroxyamino acids. Addition of a phosphate group alters the local charge on the acceptor molecule, causing internal conformational changes and potentially influencing intermolecular contacts. Reversible protein phosphorylation is the primary method for regulating protein activity in eukaryotic cells. In general, proteins are activated by phosphorylation in response to extracellular signals such as hormones, neurotransmitters, and growth and differentiation factors. The activated proteins initiate the cell's intracellular response by way of intracellular signaling pathways and second messenger molecules such as cyclic nucleotides, calcium-calmodulin, inositol, and various mitogens, that regulate protein phosphorylation.

[0003] Kinases are involved in all aspects of a cell's function, from basic metabolic processes, such as glycolysis, to cell-cycle regulation, differentiation, and communication with the extracellular environment through signal transduction cascades. Inappropriate phosphorylation of proteins in cells has been linked to changes in cell cycle progression and cell differentiation. Changes in the cell cycle have been linked to induction of apoptosis or cancer. Changes in cell differentiation have been linked to diseases and disorders of the reproductive system, immune system, and skeletal muscle.

[0004] There are two classes of protein kinases. One class, protein tyrosine kinases (PTKs), phosphorylates tyrosine residues, and the other class, protein serine/threonine kinases (STKs), phosphorylates serine and threonine residues. Some PTKs and STKs possess structural characteristics of both families and have dual specificity for both tyrosine and serine/threonine residues. Almost all kinases contain a conserved 250-300 amino acid catalytic domain containing specific residues and sequence motifs characteristic of the kinase family. The protein kinase catalytic domain can be further divided into 11 subdomains. N-terminal subdomains I-IV fold into a two-lobed structure which binds and orients the ATP donor molecule, and subdomain V spans the two lobes. C-terminal subdomains VI-XI bind the protein substrate and transfer the gamma phosphate from ATP to the hydroxyl group of a tyrosine, serine, or threonine residue. Each of the II subdomains contains specific catalytic residues or amino acid motifs characteristic of that subdomain. For example, subdomain I contains an 8-amino acid glycine-rich ATP binding consensus motif, subdomain II contains a critical lysine residue required for maximal catalytic activity, and subdomains VI through IX comprise the highly conserved catalytic core. PTKs and STKs also contain distinct sequence motifs in subdomains VI and VIII which may confer hydroxyamino acid specificity.

[0005] In addition, kinases may also be classified by additional amino acid sequences, generally between 5 and 100 residues, which either flank or occur within the kinase domain. These additional amino acid sequences regulate kinase activity and determine substrate specificity. (Reviewed in Hardie, G. and S. Hanks (1995) The Protein Kinase Facts Book, Vol I, pp. 17-20 Academic Press, San Diego Calif.). In particular, two protein kinase signature sequences have been identified in the kinase domain, the first containing an active site lysine residue involved in ATP binding, and the second containing an aspartate residue important for catalytic activity. If a protein analyzed includes the two protein kinase signatures, the probability of that protein being a protein kinase is close to 100% (PROSRM: PDOC00100, November 1995).

[0006] Protein Tyrosine Kinases

[0007] Protein tyrosine kinases (PTKs) may be classified as either transmembrane, receptor PTKs or nontransmembrane, nonreceptor PTK proteins. Transmembrane tyrosine kinases function as receptors for most growth factors. Growth factors bind to the receptor tyrosine kinase (RTK), which causes the receptor to phosphorylate itself (autophosphorylation) and specific intracellular second messenger proteins. Growth factors (GF) that associate with receptor PTKs include epidermal GF, platelet-derived GF, fibroblast GF, hepatocyte GF, insulin and insulinike GFs, nerve GF, vascular endothelial GF, and macrophage colony stimulating factor.

[0008] Nontransmembrane, nonreceptor PTKs lack transmembrane regions and, instead, form signaling complexes with the cytosolic domains of plasma membrane receptors. Receptors that function through non-receptor PTKs include those for cytokines and hormones (growth hormone and prolactin), and antigen-specific receptors on T and B lymphocytes.

[0009] Many PTKs were first identified as oncogene products in cancer cells in which PTK activation was no longer subject to normal cellular controls. In fact, about one third of the known oncogenes encode PTKs. Furthermore, cellular transformation (oncogenesis) is often accompanied by increased tyrosine phosphorylation activity (Charbonneau, H. and N. K. Tonks (1992) Annu. Rev. Cell Biol. 8:463493). Regulation of PTK activity may therefore be an important strategy in controlling some types of cancer.

[0010] Protein Serine/Threonine Kinases

[0011] Protein serine/threonine kinases (STKs) are nontransmembrane proteins. A subclass of STKs are known as ERKs (extracellular signal regulated kinases) or MAPs (mitogen-activated protein kinases) and are activated after cell stimulation by a variety of hormones and growth factors. Cell stimulation induces a signaling cascade leading to phosphorylation of MEK (MAPf/ERK kinase) which, in turn, activates ERK via serine and threonine phosphorylation. A varied number of proteins represent the downstream effectors for the active ERK and implicate it in the control of cell proliferation and differentiation, as well as regulation of the cytoskeleton. Activation of ERK is normally transient, and cells possess dual specificity phosphatases that are responsible for its down-regulation. Also, numerous studies have shown that elevated ERK activity is associated with some cancers. Other STKs include the second messenger dependent protein kinases such as the cyclic-AMP dependent protein kinases (PKA), calcium-calmodulin (CaM) dependent protein kinases, and the mitogen-activated protein kinases (MAP); the cyclin-dependent protein linases; checkpoint and cell cycle kinases; Numb-associated kinase (Nak); human Fused (hFu); proliferation-related kinases; 5'-AMP-activated protein kinases; and kinases involved in apoptosis.

[0012] The second messenger dependent protein kinases primarily mediate the effects of second messengers such as cyclic AMP (cAMP), cyclic GMP, inositol triphosphate, phosphatidylinositol, 3,4,5-triphosphate, cyclic ADP ribose, arachidonic acid, diacylglycerol and calcium-calmodulin. The PKAs are involved in mediating hormone-induced cellular responses and are activated by cAMP produced within the cell in response to hormone stimulation. cAMP is an intracellular mediator of hormone action in all animal cells that have been studied. Hormone-induced cellular responses include thyroid hormone secretion, cortisol secretion, progesterone secretion, glycogen breakdown, bone resorption, and regulation of heart rate and force of heart muscle contraction. PKA is found in all animal cells and is thought to account for the effects of cAMP in most of these cells. Altered PKA expression is implicated in a variety of disorders and diseases including cancer, thyroid disorders, diabetes, atherosclerosis, and cardiovascular disease (Isselbacher, K. J. et al. (1994) Harrison's Principles of Internal Medicine, McGraw-Hill, New York N.Y., pp. 416-431, 1887).

[0013] The casein kinase I (CKI) gene family is another subfamily of serine/threonine protein kinases. This continuously expanding group of kinases have been implicated in the regulation of numerous cytoplasmic and nuclear processes, including cell metabolism, and DNA replication and repair. CKI enzymes are present in the membranes, nucleus, cytoplasm and cytoskeleton of eukaryotic cells, and on the mitotic spindles of mammalian cells (Fish, K. J. et al. (1995) J. Biol. Chem. 270:14875-14883).

[0014] The CKI family members all have a short amino-terminal domain of 9-76 amino acids, a highly conserved kinase domain of 284 amino acids, and a variable carboxyl-terminal domain that ranges from 24 to over 200 amino acids in length (Cegielska, A. et al. (1998) J. Biol. Chem. 273:1357-1364). The CKI family is comprised of highly related proteins, as seen by the identification of isoforms of casein kinase I from a variety of sources. There are at least five mammalian isoforms, .alpha., .beta., .gamma., .delta., and .epsilon.. Fish et al., identified CKI-epsilon from a human placenta cDNA library. It is a basic protein of 416 amino acids and is closest to CKI-delta. Through recombinant expression, it was determined to phosphorylate known CKI substrates and was inhibited by the CKI-specific inhibitor CKI-7. The human gene for CKI-epsilon was able to rescue yeast with a slow-growth phenotype caused by deletion of the yeast CKI locus, HRR250 (Fish et al., supra).

[0015] The mammalian circadian mutation tau was found to be a semidominant autosomal allele of CKI-epsilon that markedly shortens period length of circadian rhythms in Syrian hamsters. The tau locus is encoded by casein kinase I-epsilon, which is also a homolog of the Drosophila circadian gene double-time. Studies of both the wildtype and tau mutant CKI-epsilon enzyme indicated that the mutant enzyme has a noticeable reduction in the maximum velocity and autophosphorylation state. Further, in vitro, CKI-epsilon is able to interact with mammalian PERIOD proteins, while the mutant enzyme is deficient in its ability to phosphorylate PERIOD. Lowrey et al., have proposed that CKI-epsilon plays a major role in delaying the negative feedback signal within the transcription-translation-based autoregulatory loop that composes the core of the circadian mechanism. Therefore the CKI-epsilon enzyme is an ideal target for pharmaceutical compounds influencing circadian rhythms, jet-lag and sleep, in addition to other physiologic and metabolic processes under circadian regulation (Lowrey, P. L. et al. (2000) Science 288:483491).

[0016] Homeodomain-interacting protein kinases (HWKs) are serine/threonine kinases and novel members of the DYRK kinase subfamily (Hofmann, T. G. et al., (2000) Biochimie 82:1123-7). HIPKs contain a conserved protein kinase domain separated from a domain that interacts with homeoproteins. HIPKs are nuclear kinases, and HIPK2 is highly expressed in neuronal tissue (Kim, Y. H. et al., (1998) J. Biol. Chem. 273:25875-9; Wang, Y. et al., (2001) Biochim. Biophys. Acta 1518:168-172). HIPKs act as corepressors for homeodomian transcription factors. This corepressor activity is seen in posttranslational modifications such as ubiquitination and phosphorylation, each are important in the regulation of cellular protein function (Kim, Y. H. et al., (1999) Proc. Nat. Acad. Sci. U.S.A. 96:123505).

[0017] Calcium-Calmodulin Dependent Protein Kinases

[0018] Calcium-calmodulin dependent (CaM) kinases are involved in regulation of smooth muscle contraction, glycogen breakdown (phosphorylase kinase), and neurotransmission (CaM kinase I and CaM kinase II). CaM dependent protein kinases are activated by calmodulin, an intracellular calcium receptor, in response to the concentration of free calcium in the cell. Many CaM kinases are also activated by phosphorylation. Some CaM kinases are also activated by autophosphorylation or by other regulatory kinases. CaM kinase I phosphorylates a variety of substrates including the neurotransmitter-related proteins synapsin I and II, the gene transcription regulator, CREB, and the cystic fibrosis conductance regulator protein, CFIR (Haribabu, B. et al. (1995) EMBO J. 14:3679-3686). CaM kinase II also phosphorylates synapsin at different sites and controls the synthesis of catecholamines in the brain through phosphorylation and activation of tyrosine hydroxylase. CaM kinase II controls the synthesis of catecholamines and seratonin, through phosphorylation/activation of tyrosine hydroxylase and tryptophan hydroxylase, respectively (Fujisawa, H. (1990) BioEssays 12:27-29). The mRNA encoding a calmodulin-binding protein kinase-like protein was found to be enriched in mammalian forebrain. This protein is associated with vesicles in both axons and dendrites and accumulates largely postnatally. The amino acid sequence of this protein is similar to CaM-dependent STKs, and the protein binds calmodulin in the presence of calcium (Godbout, M. et al. (1994) J. Neurosci. 14:1-13).

[0019] Mitogen-Activated Protein Kinases

[0020] The mitogen-activated protein kinases (MAP) which mediate signal transduction from the cell surface to the nucleus via phosphorylation cascades are another STK family that regulates intracellular signaling pathways. Several subgroups have been identified, and each manifests different substrate specificities and responds to distinct extracellular stimuli (Egan, S. E. and R. A. Weinberg (1993) Nature 365:781-783). MAP kinase signaling pathways are present in mammalian cells as well as in yeast. The extracellular stimuli which activate MAP kinase pathways include epidermal growth factor (EGF), ultraviolet light, hyperosmolar medium, heat shock, endotoxic lipopolysaccharide (LPS), and pro-inflammatory cytokines such as tumor necrosis factor (TNF) and interleukin-1 (IL-1). Altered MAP kinase expression is implicated in a variety of disease conditions including cancer, inflammation, immune disorders, and disorders affecting growth and development.

[0021] Cyclin-Dependent Protein Kinases

[0022] The cyclin-dependent protein kinases (CDKs) are STKs that control the progression of cells through the cell cycle. The entry and exit of a cell from mitosis are regulated by the synthesis and destruction of a family of activating proteins called cyclins. Cyclins are small regulatory proteins that bind to and activate CDKs, which then phosphorylate and activate selected proteins involved in the mitotic process. CDKs are unique in that they require multiple inputs to become activated. In addition to cyclin binding, CDK activation requires the phosphorylation of a specific threonine residue and the dephosphorylation of a specific tyrosine residue on the CDK.

[0023] Another family of STKs associated with the cell cycle are the NIMA (never in mitosis)-related kinases (Neks). Both CDKs and Neks are involved in duplication, maturation, and separation of the microtubule organizing center, the centrosome, in animal cells (Fry, A. M. et al. (1998) EMBO J. 17:470481).

[0024] Checkpoint and Cell Cycle Kinases

[0025] In the process of cell division, the order and timing of cell cycle transitions are under control of cell cycle checkpoints, which ensure that critical events such as DNA replication and chromosome segregation are carried out with precision. If DNA is damaged, e.g. by radiation, a checkpoint pathway is activated that arrests the cell cycle to provide time for repair. If the damage is extensive, apoptosis is induced. In the absence of such checkpoints, the damaged DNA is inherited by aberrant cells which may cause proliferative disorders such as cancer. Protein kinases play an important role in this process. For example, a specific kinase, checkpoint kinase 1 (Chk1), has been identified in yeast and mammals, and is activated by DNA damage in yeast. Activation of Chk1 leads to the arrest of the cell at the G2/A transition (Sanchez, Y. et al. (1997) Science 277:1497-1501). Specifically, Chk1 phosphorylates the cell division cycle phosphatase CDC25, inhibiting its normal function which is to dephosphorylate and activate the cyclin-dependent kinase Cdc2. Cdc2 activation controls the entry of cells into mitosis (Peng, C. -Y. et al. (1997) Science 277:1501-1505). Thus, activation of Chk1 prevents the damaged cell from entering mitosis. A similar deficiency in a checkpoint kinase, such as Chk1, may also contribute to cancer by failure to arrest cells with damaged DNA at other checkpoints such as G2/M.

[0026] Proliferation-Related Kinases

[0027] Proliferation-related kinase is a serum/cytokine inducible STK that is involved in regulation of the cell cycle and cell proliferation in human megakarocytic cells (Li, B. et al. (1996) J. Biol. Chem. 271:19402-19408). Proliferation-related kinase is related to the polo (derived from Drosophila polo gene) family of STKs implicated in cell division. Proliferation-related kinase is downregulated in lung tumor tissue and may be a proto-oncogene whose deregulated expression in normal tissue leads to oncogenic transformation.

[0028] 5'-AMP-Activated Protein Kinase

[0029] A ligand-activated STK protein kinase is 5'-AMP-activated protein kinase (AMPK) (Gao, G. et al. (1996) J. Biol. Chem. 271:8675-8681). Mammalian AMPK is a regulator of fatty acid and sterol synthesis through phosphorylation of the enzymes acetyl-CoA carboxylase and hydroxymethylglutaryl-CoA reductase and mediates responses of these pathways to cellular stresses such as heat shock and depletion of glucose and ATP. AMPK is a heterotrimeric complex comprised of a catalytic alpha subunit and two non-catalytic beta and gamma subunits that are believed to regulate the activity of the alpha subunit. Subunits of AMPK have a much wider distribution in non-lipogenic tissues such as brain, heart, spleen, and lung than expected. This distribution suggests that its role may extend beyond regulation of lipid metabolism alone.

[0030] Kinases in Apoptosis

[0031] Apoptosis is a highly regulated signaling pathway leading to cell death that plays a crucial role in tissue development and homeostasis. Deregulation of this process is associated with the pathogenesis of a number of diseases including autoimmune disease, neurodegenerative disorders, and cancer. Various STKs play key roles in this process. ZIP kinase is an STK containing a C-terminal leucine zipper domain in addition to its N-terminal protein kinase domain. This C-terminal domain appears to mediate homodimerization and activation of the kinase as well as interactions with transcription factors such as activating transcription factor, ATF4, a member of the cyclic-AMP responsive element binding protein (AT/CREB) family of transcriptional factors (Sanjo, H. et al. (1998) J. Biol. Chem. 273:29066-29071). DRAK1 and DRAK2 are STKs that share homology with the death-associated protein kinases (DAP kinases), known to function in interferon-.gamma. induced apoptosis (Sanjo et al., supra). Like ZIP kinase, DAP kinases contain a C-terminal protein-protein interaction domain, in the form of ankyrin repeats, in addition to the N-terminal kinase domain. ZIP, DAP, and DRAK kinases induce morphological changes associated with apoptosis when transfected into NIH3T3 cells (Sanjo et al., supra). However, deletion of either the N-terminal kinase catalytic domain or the C-terminal domain of these proteins abolishes apoptosis activity, indicating that in addition to the kinase activity, activity in the C-terminal domain is also necessary for apoptosis, possibly as an interacting domain with a regulator or a specific substrate.

[0032] RICK is another STK recently identified as mediating a specific apoptotic pathway involving the death receptor, CD95 (Inohara, N. et al. (1998) J. Biol. Chem. 273:12296-12300). CD95 is a member of the tumor necrosis factor receptor superfamily and plays a critical role in the regulation and homeostasis of the immune system (Nagata, S. (1997) Cell 88:355-365). The CD95 receptor signaling pathway involves recruitment of various intracellular molecules to a receptor complex following ligand binding. This process includes recruitment of the cysteine protease caspase-8 which, in turn, activates a caspase cascade leading to cell death. RICK is composed of an N-terminal kinase catalytic domain and a C-terminal "caspase-recruitment" domain that interacts with caspase-like domains, indicating that RICK plays a role in the recruitment of caspase-8. This interpretation is supported by the fact that the expression of RICK in human 293T cells promotes activation of caspase-8 and potentiates the induction of apoptosis by various proteins involved in the CD95 apoptosis pathway (Inohara et al., supra).

[0033] Mitochondrial Protein Kinases

[0034] A novel class of eukaryotic kinases, related by sequence to prokaryotic histidine protein kinases, are the mitochondrial protein kinases (MPKs) which seem to have no sequence similarity with other eukaryotic protein kinases. These protein kinases are located exclusively in the mitochondrial matrix space and may have evolved from genes originally present in respiration-dependent bacteria which were endocytosed by primitive eukaryotic cells. MPKs are responsible for phospborylation and inactivation of the branched-chain alpha-ketoacid dehydrogenase and pyruvate dehydrogenase complexes (Harris, R. A. et al. (1995) Adv. Enzyme Regul. 34:147-162). Five MPKs have been identified. Four members correspond to pyruvate dehydrogenase kinase isozymes, regulating the activity of the pyruvate dehydrogenase complex, which is an important regulatory enzyme at the interface between glycolysis and the citric acid cycle. The fifth member corresponds to a branched-chain alpha-ketoacid dehydrogenase kinase, important in the regulation of the pathway for the disposal of branched-chain amino acids. (Harris, R. A. et al. (1997) Adv. Enzyme Regul. 37:271-293). Both starvation and the diabetic state are known to result in a great increase in the activity of the pyruvate dehydrogenase kinase in the liver, heart and muscle of the rat. This increase contributes in both disease states to the phosphorylation and inactivation of the pyruvate dehydrogenase complex and conservation of pyruvate and lactate for gluconeogenesis (Harris (1995) supra).

[0035] Kinases with Non-Protein Substrates

[0036] Lipid and Inositol kinases

[0037] Lipid kinases phosphorylate hydroxyl residues on lipid head groups. A family of kinases involved in phosphorylation of phosphatidylinositol (PI) has been described, each member phosphorylating a specific carbon on the inositol ring (Leevers, S. J. et al. (1999) Curr. Opin. Cell. Biol. 11:219-225). The phosphorylation of phosphatidylinositol is involved in activation of the protein kinase C signaling pathway. The inositol phospholipids (phosphoinositides) intracellular signaling pathway begins with binding of a signaling molecule to a G-protein linked receptor in the plasma membrane. This leads to the phosphorylation of phosphatidylinositol (PI) residues on the inner side of the plasma membrane by inositol kinases, thus converting PI residues to the biphosphate state (PIP.sub.2). PIP.sub.2 is then cleaved into inositol triphosphate (IP.sub.3) and diacylglycerol. These two products act as mediators for separate signaling pathways. Cellular responses that are mediated by these pathways are glycogen breakdown in the liver in response to vasopressin, smooth muscle contraction in response to acetylcholine, and thrombin-induced platelet aggregation.

[0038] PI 3-kinase (PI3K), which phosphorylates the D3 position of PI and its derivatives, has a central role in growth factor signal cascades involved in cell growth, differentiation, and metabolism. PI3K is a heterodimer consisting of an adapter subunit and a catalytic subunit. The adapter subunit acts as a scaffolding protein, interacting with specific tyrosine-phosphorylated proteins, lipid moieties, and other cytosolic factors. When the adapter subunit binds tyrosine phosphorylated targets, such as the insulin responsive substrate (IRS)-1, the catalytic subunit is activated and converts PI (4,5) bisphosphate (PIP.sub.2) to PI (3,4,5) P.sub.3 (PIP.sub.3). PIP.sub.3 then activates a number of other proteins, including PKA, protein kinase B (PKB), protein kinase C (PKC), glycogen synthase kinase (GSK)-3, and p70 ribosomal s6 kinase. PI3K also interacts directly with the cytoskeletal organizing proteins, Rac, rho, and cdc42 (Shepherd, P. R. et al. (1998) Biochem. J. 333:471490). Animal models for diabetes, such as obese and fat mice, have altered PI3K adapter subunit levels. Specific mutations in the adapter subunit have also been found in an insulin-resistant Danish population, suggesting a role for PI3K in type-2 diabetes (Shepard, supra).

[0039] An example of lipid kinase phosphorylation activity is the phosphorylation of D-erythro-sphingosine to the sphingolipid metabolite, sphingosine-1-phosphate (SPP). SPP has emerged as a novel lipid second-messenger with both extracellular and intracellular actions (Kohama, T. et al. (1998) J. Biol. Chem. 273:23722-23728). Extracellularly, SPP is a ligand for the G-protein coupled receptor EDG-1 (endothelial-derived, G-protein coupled receptor). Intracellularly, SPP regulates cell growth, survival, motility, and cytoskeletal changes. SPP levels are regulated by sphingosine kinases that specifically phosphorylate D-erythro-sphingosine to SPP. The importance of sphingosine kinase in cell signaling is indicated by the fact that various stimuli, including platelet-derived growth factor (PDGF), nerve growth factor, and activation of protein kinase C, increase cellular levels of SPP by activation of sphingosine kinase, and the fact that competitive inhibitors of the enzyme selectively inhibit cell proliferation induced by PDGF (Kohama et al., supra).

[0040] Purine Nucleotide Kinases

[0041] The purine nucleotide kinases, adenylate kinase (ATP:AMP phosphotransferase, or AdK) and guanylate kinase (ATP:GMP phosphotransferase, or GuK) play a key role in nucleotide metabolism and are crucial to the synthesis and regulation of cellular levels of ATP and GTP, respectively. These two molecules are precursors in DNA and RNA synthesis in growing cells and provide the primary source of biochemical energy in cells (ATP), and signal transduction pathways (GTP). Inhibition of various steps in the synthesis of these two molecules has been the basis of many antiproliferative drugs for cancer and antiviral therapy (Pillwein, K. et al. (1990) Cancer Res. 50:1576-1579).

[0042] AdK is found in almost all cell types and is especially abundant in cells having high rates of ATP synthesis and utilization such as skeletal muscle. In these cells AdK is physically associated with mitochondria and myofibrils, the subcellular structures that are involved in energy production and utilization, respectively. Recent studies have demonstrated a major function for AdK in transferring high energy phosphoryls from metabolic processes generating ATP to cellular components consuming ATP (Zeleznikar, R. J. et al. (1995) J. Biol. Chem. 270:7311-7319). Thus AdK may have a pivotal role in maintaining energy production in cells, particularly those having a high rate of growth or metabolism such as cancer cells, and may provide a target for suppression of its activity to treat certain cancers. Alternatively, reduced AdK activity may be a source of various metabolic, muscle-energy disorders that can result in cardiac or respiratory failure and may be treatable by increasing AdK activity.

[0043] GuK, in addition to providing a key step in the synthesis of GTP for RNA and DNA synthesis, also fulfills an essential function in signal transduction pathways of cells through the regulation of GDP and GTP. Specifically, GTP binding to membrane associated G proteins mediates the activation of cell receptors, subsequent intracellular activation of adenyl cyclase, and production of the second messenger, cyclic AMP. GDP binding to G proteins inhibits these processes. GDP and GTP levels also control the activity of certain oncogenic proteins such as p21.sup.ras known to be involved in control of cell proliferation and oncogenesis (Bos, J. L. (1989) Cancer Res. 49:46824689). High ratios of GTP:GDP caused by suppression of GuK cause activation of p21.sup.ras and promote oncogenesis. Increasing GuK activity to increase levels of GDP and reduce the GTP:GDP ratio may provide a therapeutic strategy to reverse oncogenesis.

[0044] GuK is an important enzyme in the phosphorylation and activation of certain antiviral drugs useful in the treatment of herpes virus infections. These drugs include the guanine homologs acyclovir and buciclovir (Miller, W. H. and R. L. Miller (1980) J. Biol. Chem. 255:7204-7207; Stenberg, K. et al. (1986) J. Biol. Chem. 261:2134-2139). Increasing GuK activity in infected cells may provide a therapeutic strategy for augmenting the effectiveness of these drugs and possibly for reducing the necessary dosages of the drugs.

[0045] Pyrimidine Kinases

[0046] The pyrimidine kinases are deoxycytidine kinase and thymidine kinase 1 and 2. Deoxycytidine kinase is located in the nucleus, and thymidine kinase 1 and 2 are found in the cytosol (Johansson, M. et al. (1997) Proc. Natl. Acad. Sci. USA 94:11941-11945). Phosphorylation of deoxyribonucleosides by pyrimidine kinases provides an alternative pathway for de novo synthesis of DNA precursors. The role of pyrimidine kinases, like purine kinases, in phosphorylation is critical to the activation of several chemotherapeutically important nucleoside analogues (Arner E. S. and S. Eriksson (1995) Pharmacol. Ther. 67:155-186).

[0047] The discovery of new human kinases and the polynucleotides encoding them satisfies a need in the art by providing new compositions which are useful in the diagnosis, prevention, and treatment of cancer, immune disorders, disorders affecting growth and development, cardiovascular diseases, and lipid disorders, and in the assessment of the effects of exogenous compounds on the expression of nucleic acid and amino acid sequences of human kinases.

SUMMARY OF THE INVNTION

[0048] The invention features purified polypeptides, human kinases, referred to collectively as "PKIN" and individually as "PKIN-1," "PKIN-2," "PKIN-3," "PKIN-4," "PKIN-5," "PKIN-6," "PKN-7," "PKIN-8," "PKIN-9," "PKIN-10," "PKIN-11," "PKIN-12," "PKI-13," "PKIN-14," "PKIN-15," "PKIN-16," "PKIN-17," and "PKIN-18." In one aspect, the invention provides an isolated polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, b) a naturally occurring polypeptide comprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18. In one alternative, the invention provides an isolated polypeptide comprising the amino acid sequence of SEQ ID NO:1-18.

[0049] The invention further provides an isolated polynucleotide encoding a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, b) a naturally occurring polypeptide comprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18. In one alternative, the polynucleotide encodes a polypeptide selected from the group consisting of SEQ ID NO:1-18. In another alternative, the polynucleotide is selected from the group consisting of SEQ ID NO:19-36.

[0050] Additionally, the invention provides a recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide encoding a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, b) a naturally occurring polypeptide comprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18. In one alternative, the invention provides a cell transformed with the recombinant polynucleotide. In another alternative, the invention provides a transgenic organism comprising the recombinant polynucleotide.

[0051] The invention also provides a method for producing a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, b) a naturally occurring polypeptide comprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18. The method comprises a) culturing a cell under conditions suitable for expression of the polypeptide, wherein said cell is transformed with a recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide encoding the polypeptide, and b) recovering the polypeptide so expressed.

[0052] Additionally, the invention provides an isolated antibody which specifically binds to a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, b) a naturally occurring polypeptide comprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18.

[0053] The invention further provides an isolated polynucleotide selected from the group consisting of a) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:19-36, b) a naturally occurring polynucleotide comprising a polynucleotide sequence at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NO:19-36, c) a polynucleotide complementary to the polynucleotide of a), d) a polynucleotide complementary to the polynucleotide of b), and e) an RNA equivalent of a)-d). In one alternative, the polynucleotide comprises at least 60 contiguous nucleotides.

[0054] Additionally, the invention provides a method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide selected from the group consisting of a) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ E) NO:19-36, b) a naturally occurring polynucleotide comprising a polynucleotide sequence at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NO:19-36, c) a polynucleotide complementary to the polynucleotide of a), d) a polynucleotide complementary to the polynucleotide of b), and e) an RNA equivalent of a)-d). The method comprises a) hybridizing the sample with a probe comprising at least 20 contiguous nucleotides comprising a sequence complementary to said target polynucleotide in the sample, and which probe specifically hybridizes to said target polynucleotide, under conditions whereby a hybridization complex is formed between said probe and said target polynucleotide or fragments thereof, and b) detecting the presence or absence of said hybridization complex, and optionally, if present, the amount thereof. In one alternative, the probe comprises at least 60 contiguous nucleotides.

[0055] The invention further provides a method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide selected from the group consisting of a) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:19-36, b) a naturally occurring polynucleotide comprising a polynucleotide sequence at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NO:19-36, c) a polynucleotide complementary to the polynucleotide of a), d) a polynucleotide complementary to the polynucleotide of b), and e) an RNA equivalent of a)-d). The method comprises a) amplifying said target polynucleotide or fragment thereof using polymerase chain reaction amplification, and b) detecting the presence or absence of said amplified target polynucleotide or fragment thereof, and, optionally, if present, the amount thereof.

[0056] The invention further provides a composition comprising an effective amount of a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, b) a naturally occurring polypeptide comprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, and a pharmaceutically acceptable excipient. In one embodiment, the composition comprises an amino acid sequence selected from the group consisting of SEQ ID NO:1-18. The invention additionally provides a method of treating a disease or condition associated with decreased expression of functional PKIN, comprising administering to a patient in need of such treatment the composition.

[0057] The invention also provides a method for screening a compound for effectiveness as an agonist of a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, b) a naturally occurring polypeptide comprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18. The method comprises a) exposing a sample comprising the polypeptide to a compound, and b) detecting agonist activity in the sample. In one alternative, the invention provides a composition comprising an agonist compound identified by the method and a pharmaceutically acceptable excipient. In another alternative, the invention provides a method of treating a disease or condition associated with decreased expression of functional PKIN, comprising administering to a patient in need of such treatment the composition.

[0058] Additionally, the invention provides a method for screening a compound for effectiveness as an antagonist of a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, b) a naturally occurring polypeptide comprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18. The method comprises a) exposing a sample comprising the polypeptide to a compound, and b) detecting antagonist activity in the sample. In one alternative, the invention provides a composition comprising an antagonist compound identified by the method and a pharmaceutically acceptable excipient. In another alternative, the invention provides a method of treating a disease or condition associated with overexpression of functional PKIN, comprising administering to a patient in need of such treatment the composition.

[0059] The invention further provides a method of screening for a compound that specifically binds to a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, b) a naturally occurring polypeptide cmoprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18. The method comprises a) combining the polypeptide with at least one test compound under suitable conditions, and b) detecting binding of the polypeptide to the test compound, thereby identifying a compound that specifically binds to the polypeptide.

[0060] The invention further provides a method of screening for a compound that modulates the activity of a polypeptide selected from the group consisting of a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, b) a naturally occurring polypeptide comprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18. The method comprises a) combining the polypeptide with at least one test compound under conditions permissive for the activity of the polypeptide, b) assessing the activity of the polypeptide in the presence of the test compound, and c) comparing the activity of the polypeptide in the presence of the test compound with the activity of the polypeptide in the absence of the test compound, wherein a change in the activity of the polypeptide in the presence of the test compound is indicative of a compound that modulates the activity of the polypeptide.

[0061] The invention further provides a method for screening a compound for effectiveness in altering expression of a target polynucleotide, wherein said target polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO:19-36, the method comprising a) exposing a sample comprising the target polynucleotide to a compound, and b) detecting altered expression of the target polynucleotide.

[0062] The invention further provides a method for assessing toxicity of a test compound, said method comprising a) treating a biological sample containing nucleic acids with the test compound; b) hybridizing the nucleic acids of the treated biological sample with a probe comprising at least 20 contiguous nucleotides of a polynucleotide selected from the group consisting of i) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:19-36, ii) a naturally occurring polynucleotide comprising a polynucleotide sequence at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NO:19-36, iii) a polynucleotide having a sequence complementary to i), iv) a polynucleotide complementary to the polynucleotide of ii), and v) an RNA equivalent of i)-iv). Hybridization occurs under conditions whereby a specific hybridization complex is formed between said probe and a target polynucleotide in the biological sample, said target polynucleotide selected from the group consisting of i) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:19-36, ii) a naturally occurring polynucleotide comprising a polynucleotide sequence at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NO:19-36, iii) a polynucleotide complementary to the polynucleotide of i), iv) a polynucleotide complementary to the polynucleotide of ii), and v) an RNA equivalent of i)-iv). Alternatively, the target polynucleotide comprises a fragment of a polynucleotide sequence selected from the group consisting of i)-v) above; c) quantifying the amount of hybridization complex; and d) comparing the amount of hybridization complex in the treated biological sample with the amount of hybridization complex in an untreated biological sample, wherein a difference in the amount of hybridization complex in the treated biological sample is indicative of toxicity of the test compound.

BRIEF DESCRIPTION OF THE TABLES

[0063] Table 1 summarizes the nomenclature for the full length polynucleotide and polypeptide sequences of the present invention.

[0064] Table 2 shows the GenBank identification number and annotation of the nearest GenBank homolog for polypeptides of the invention. The probability score for the match between each polypeptide and its GenBank homolog is also shown.

[0065] Table 3 shows structural features of polypeptide sequences of the invention, including predicted motifs and domains, along with the methods, algorithms, and searchable databases used for analysis of the polypeptides.

[0066] Table 4 lists the cDNA and genomic DNA fragments which were used to assemble polynucleotide sequences of the invention, along with selected fragments of the polynucleotide sequences.

[0067] Table 5 shows the representative cDNA library for polynucleotides of the invention.

[0068] Table 6 provides an appendix which describes the tissues and vectors used for construction of the cDNA libraries shown in Table 5.

[0069] Table 7 shows the tools, programs, and algorithms used to analyze the polynucleotides and polypeptides of the invention, along with applicable descriptions, references, and threshold parameters.

DESCRIPTION OF THE INVENTION

[0070] Before the present proteins, nucleotide sequences, and methods are described, it is understood that this invention is not limited to the particular machines, materials and methods described, as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention which will be limited only by the appended claims.

[0071] It must be noted that as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, a reference to "a host cell" includes a plurality of such host cells, and a reference to "an antibody" is a reference to one or more antibodies and equivalents thereof known to those skilled in the art, and so forth.

[0072] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any machines, materials, and methods similar or equivalent to those described herein can be used to practice or test the present invention, the preferred machines, materials and methods are now described. All publications mentioned herein are cited for the purpose of describing and disclosing the cell lines, protocols, reagents and vectors which are reported in the publications and which might be used in connection with the invention. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

[0073] Definitions

[0074] "PKIN" refers to the amino acid sequences of substantially purified PKIN obtained from any species, particularly a mammalian species, including bovine, ovine, porcine, murine, equine, and human, and from any source, whether natural, synthetic, semi-synthetic, or recombinant.

[0075] The term "agonist" refers to a molecule which intensifies or mimics the biological activity of PKIN. Agonists may include proteins, nucleic acids, carbohydrates, small molecules, or any other compound or composition which modulates the activity of PKIN either by directly interacting with PKIN or by acting on components of the biological pathway in which PKIN participates.

[0076] An "allelic variant" is an alternative form of the gene encoding PKEN. Allelic variants may result from at least one mutation in the nucleic acid sequence and may result in altered mRNAs or in polypeptides whose structure or function may or may not be altered. A gene may have none, one, or many allelic variants of its naturally occurring form. Common mutational changes which give rise to allelic variants are generally ascribed to natural deletions, additions, or substitutions of nucleotides. Each of these types of changes may occur alone, or in combination with the others, one or more times in a given sequence.

[0077] "Altered" nucleic acid sequences encoding PKIN include those sequences with deletions, insertions, or substitutions of different nucleotides, resulting in a polypeptide the same as PKIN or a polypeptide with at least one functional characteristic of PKIN. Included within this definition are polymorphisms which may or may not be readily detectable using a particular oligonucleotide probe of the polynucleotide encoding PKIN, and improper or unexpected hybridization to allelic variants, with a locus other than the normal chromosomal locus for the polynucleotide sequence encoding PKIN. The encoded protein may also be "altered," and may contain deletions, insertions, or substitutions of amino acid residues which produce a silent change and result in a functionally equivalent PKIN. Deliberate amino acid substitutions may be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and/or the amphipathic nature of the residues, as long as the biological or immunological activity of PKIN is retained. For example, negatively charged amino acids may include aspartic acid and glutamic acid, and positively charged amino acids may include lysine and arginine. Amino acids with uncharged polar side chains having similar hydrophilicity values may include: asparagine and glutamine; and serine and threonine. Amino acids with uncharged side chains having similar hydrophilicity values may include: leucine, isoleucine, and valine; glycine and alanine; and phenylalanine and tyrosine.

[0078] The terms "amino acid" and "amino acid sequence" refer to an oligopeptide, peptide, polypeptide, or protein sequence, or a fragment of any of these, and to naturally occurring or synthetic molecules. Where "amino acid sequence" is recited to refer to a sequence of a naturally occurring protein molecule, "amino acid sequence" and like terms are not meant to limit the amino acid sequence to the complete native amino acid sequence associated with the recited protein molecule.

[0079] "Amplification" relates to the production of additional copies of a nucleic acid sequence. Amplification is generally carried out using polymerase chain reaction (PCR) technologies well known in the art.

[0080] The term "antagonist" refers to a molecule which inhibits or attenuates the biological activity of PKIN. Antagonists may include proteins such as antibodies, nucleic acids, carbohydrates, small molecules, or any other compound or composition which modulates the activity of PKIN either by directly interacting with PKIN or by acting on components of the biological pathway in which PKIN participates.

[0081] The term "antibody" refers to intact immunoglobulin molecules as well as to fragments thereof, such as Fab, F(ab').sub.2, and Fv fragments, which are capable of binding an epitopic determinant. Antibodies that bind PKIN polypeptides can be prepared using intact polypeptides or using fragments containing small peptides of interest as the immunizing antigen. The polypeptide or oligopeptide used to immunize an animal (e.g., a mouse, a rat, or a rabbit) can be derived from the translation of RNA, or synthesized chemically, and can be conjugated to a carrier protein if desired. Commonly used carriers that are chemically coupled to peptides include bovine serum albumin, thyroglobulin, and keyhole limpet hemocyanin (KLH). The coupled peptide is then used to immunize the animal.

[0082] The term "antigenic determinant" refers to that region of a molecule (i.e., an epitope) that makes contact with a particular antibody. When a protein or a fragment of a protein is used to immunize a host animal, numerous regions of the protein may induce the production of antibodies which bind specifically to antigenic determinants (particular regions or three-dimensional structures on the protein). An antigenic determinant may compete with the intact antigen (i.e., the immunogen used to elicit the immune response) for binding to an antibody.

[0083] The term "antisense" refers to any composition capable of base-pairing with the "sense" (coding) strand of a specific nucleic acid sequence. Antisense compositions may include DNA; RNA; peptide nucleic acid (PNA); oligonucleotides having modified backbone linkages such as phosphorothioates, methylphosphonates, or benzylphosphonates; oligonucleotides having modified sugar groups such as 2'-methoxyethyl sugars or 2'-methoxyethoxy sugars; or oligonucleotides having modified bases such as 5-methyl cytosine, 2'-deoxyuracil, or 7-deaza-2'-deoxyguanosine. Antisense molecules may be produced by any method including chemical synthesis or transcription. Once introduced into a cell, the complementary antisense molecule base-pairs with a naturally occurring nucleic acid sequence produced by the cell to form duplexes which block either transcription or translation. The designation "negative" or "minus" can refer to the antisense strand, and the designation "positive" or "plus" can refer to the sense strand of a reference DNA molecule.

[0084] The term "biologically active" refers to a protein having structural, regulatory, or biochemical functions of a naturally occurring molecule. Likewise, "immunologically active" or "immunogenic" refers to the capability of the natural, recombinant, or synthetic PKIN, or of any oligopeptide thereof, to induce a specific immune response in appropriate animals or cells and to bind with specific antibodies.

[0085] "Complementary" describes the relationship between two single-stranded nucleic acid sequences that anneal by base-pairing. For example, 5'-AGT-3' pairs with its complement, 3'-TCA-5'.

[0086] A "composition comprising a given polynucleotide sequence" and a "composition comprising a given amino acid sequence" refer broadly to any composition containing the given polynucleotide or amino acid sequence. The composition may comprise a dry formulation or an aqueous solution. Compositions comprising polynucleotide sequences encoding PKIN or fragments of PKIN may be employed as hybridization probes. The probes may be stored in freeze-dried form and may be associated with a stabilizing agent such as a carbohydrate. In hybridizations, the probe may be deployed in an aqueous solution containing salts (e.g., NaCl), detergents (e.g., sodium dodecyl sulfate; SDS), and other components (e.g., Denhardt's solution, dry milk, salmon sperm DNA, etc.). "Consensus sequence" refers to a nucleic acid sequence which has been subjected to repeated DNA sequence analysis to resolve uncalled bases, extended using the XL-PCR kit (Applied Biosystems, Foster City Calif.) in the 5' and/or the 3' direction, and resequenced, or which has been assembled from one or more overlapping cDNA, EST, or genomic DNA fragments using a computer program for fragment assembly, such as the GELVIEW fragment assembly system (GCG, Madison Wis.) or Phrap (University of Washington, Seattle Wash.). Some sequences have been both extended and assembled to produce the consensus sequence.

[0087] "Conservative amino acid substitutions" are those substitutions that are predicted to least interfere with the properties of the original protein, i.e., the structure and especially the function of the protein is conserved and not significantly changed by such substitutions. The table below shows amino acids which may be substituted for an original amino acid in a protein and which are regarded as conservative amino acid substitutions.

1 Original Residue Conservative Substitution Ala Gly, Ser Arg His, Lys Asn Asp, Gln, His Asp Asn, Gln Cys Ala, Ser Gln Asn, Gln, His Glu Asp, Gln, His Gly Ala His Asn, Arg, Gln, Glu Ile Leu, Val Leu Ile, Val Lys Arg, Gln, Gln Met Leu, Ile Phe His, Met, Leu, Trp, Tyr Ser Cys, Thr Thr Ser, Val Trp Phe, Tyr Tyr His, Phe, Trp Val Ile, Leu, Thr

[0088] Conservative amino acid substitutions generally maintain (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a beta sheet or alpha helical conformation, (b) the charge or hydrophobicity of the molecule at the site of the substitution, and/or (c) the bulk of the side chain.

[0089] A "deletion" refers to a change in the amino acid or nucleotide sequence that results in the absence of one or more amino acid residues or nucleotides.

[0090] The term "derivative" refers to a chemically modified polynucleotide or polypeptide. Chemical modifications of a polynucleotide can include, for example, replacement of hydrogen by an alkyl, acyl, hydroxyl, or amino group. A derivative polynucleotide encodes a polypeptide which retains at least one biological or immunological function of the natural molecule. A derivative polypeptide is one modified by glycosylation, pegylation, or any similar process that retains at least one biological or immunological function of the polypeptide from which it was derived.

[0091] A "detectable label" refers to a reporter molecule or enzyme that is capable of generating a measurable signal and is covalently or noncovalently joined to a polynucleotide or polypeptide.

[0092] "Differential expression" refers to increased or upregulated; or decreased, downregulated, or absent gene or protein expression, determined by comparing at least two different samples. Such comparisons may be carried out between, for example, a treated and an untreated sample, or a diseased and a normal sample.

[0093] A "fragment" is a unique portion of PKIN or the polynucleotide encoding PKIN which is identical in sequence to but shorter in length than the parent sequence. A fragment may comprise up to the entire length of the defined sequence, minus one nucleotide/amino acid residue. For example, a fragment may comprise from 5 to 1000 contiguous nucleotides or amino acid residues. A fragment used as a probe, primer, antigen, therapeutic molecule, or for other purposes, may be at least 5, 10, 15, 16, 20, 25, 30, 40, 50, 60, 75, 100, 150, 250 or at least 500 contiguous nucleotides or amino acid residues in length. Fragments may be preferentially selected from certain regions of a molecule. For example, a polypeptide fragment may comprise a certain length of contiguous amino acids selected from the first 250 or 500 amino acids (or first 25% or 50%) of a polypeptide as shown in a certain defined sequence. Clearly these lengths are exemplary, and any length that is supported by the specification, including the Sequence Listing, tables, and figures, may be encompassed by the present embodiments.

[0094] A fragment of SEQ ID NO:19-36 comprises a region of unique polynucleotide sequence that specifically identifies SEQ ID NO:19-36, for example, as distinct from any other sequence in the genome from which the fragment was obtained. A fragment of SEQ ID NO:19-36 is useful, for example, in hybridization and amplification technologies and in analogous methods that distinguish SEQ ID NO:19-36 from related polynucleotide sequences. The precise length of a fragment of SEQ ID NO:19-36 and the region of SEQ ID NO:19-36 to which the fragment corresponds are routinely determinable by one of ordinary skill in the art based on the intended purpose for the fragment.

[0095] A fragment of SEQ ID NO:1-18 is encoded by a fragment of SEQ ID NO:19-36. A fragment of SEQ ID NO:1-18 comprises a region of unique amino acid sequence that specifically identifies SEQ ID NO:1-18. For example, a fragment of SEQ ID NO:1-18 is useful as an immunogenic peptide for the development of antibodies that specifically recognize SEQ ID NO:1-18. The precise length of a fragment of SEQ ID NO:1-18 and the region of SEQ ID NO:1-18 to which the fragment corresponds are routinely determinable by one of ordinary skill in the art based on the intended purpose for the fragment.

[0096] A "full length" polynucleotide sequence is one containing at least a translation initiation codon (e.g., methionine) followed by an open reading frame and a translation termination codon. A "full length" polynucleotide sequence encodes a "full length" polypeptide sequence.

[0097] "Homology" refers to sequence similarity or, interchangeably, sequence identity, between two or more polynucleotide sequences or two or more polypeptide sequences.

[0098] The terms "percent identity" and "% identity," as applied to polynucleotide sequences, refer to the percentage of residue matches between at least two polynucleotide sequences aligned using a standardized algorithm. Such an algorithm may insert, in a standardized and reproducible way, gaps in the sequences being compared in order to optimize alignment between two sequences, and therefore achieve a more meaningful comparison of the two sequences.

[0099] Percent identity between polynucleotide sequences may be determined using the default parameters of the CLUSTAL V algorithm as incorporated into the MEGALIGN version 3.12e sequence alignment program. This program is part of the LASERGENE software package, a suite of molecular biological analysis programs (DNASTAR, Madison Wis.). CLUSTAL V is described in Higgins, D. G. and P. M. Sharp (1989) CABIOS 5:151-153 and in Higgins, D. G. et al. (1992) CABIOS 8.189-191. For pairwise alignments of polynucleotide sequences, the default parameters are set as follows: Ktuple=2, gap penalty=5, window=4, and "diagonals saved"=4. The "weighted" residue weight table is selected as the default. Percent identity is reported by CLUSTAL V as the "percent similarity" between aligned polynucleotide sequences.

[0100] Alternatively, a suite of commonly used and freely available sequence comparison algorithms is provided by the National Center for Biotechnology Information (NCBI) Basic Local Alignment Search Tool (BLAST) (Altschul, S. F. et al. (1990) J. Mol. Biol. 215:403410), which is available from several sources, including the NCBL Bethesda, Md., and on the Internet at http://www.ncbi.nlm.nih.gov/BLAST/. The BLAST software suite includes various sequence analysis programs including "blastn," that is used to align a known polynucleotide sequence with other polynucleotide sequences from a variety of databases. Also available is a tool called "BLAST 2 Sequences" that is used for direct pairwise comparison of two nucleotide sequences. "BLAST 2 Sequences" can be accessed and used interactively at http://www.ncbi.nlm.nih.gov/gorf/b12.h- tml. The "BLAST 2 Sequences" tool can be used for both blastn and blastp (discussed below). BLAST programs are commonly used with gap and other parameters set to default settings. For example, to compare two nucleotide sequences, one may use blastn with the "BLAST 2 Sequences" tool Version 2.0.12 (April-21-2000) set at default parameters. Such default parameters may be, for example:

[0101] Matrix: BLOSUM62

[0102] Reward for match: 1

[0103] Penalty for mismatch: -2

[0104] Open Gap: 5 and Extension Gap: 2 penalties

[0105] Gap x drop-off 50

[0106] Expect: 10

[0107] Word Size: 11

[0108] Filter: on

[0109] Percent identity may be measured over the length of an entire defined sequence, for example, as defined by a particular SEQ ID number, or may be measured over a shorter length, for example, over the length of a fragment taken from a larger, defined sequence, for instance, a fragment of at least 20, at least 30, at least 40, at least 50, at least 70, at least 100, or at least 200 contiguous nucleotides. Such lengths are exemplary only, and it is understood that any fragment length supported by the sequences shown herein, in the tables, figures, or Sequence Listing, may be used to describe a length over which percentage identity may be measured.

[0110] Nucleic acid sequences that do not show a high degree of identity may nevertheless encode similar amino acid sequences due to the degeneracy of the genetic code. It is understood that changes in a nucleic acid sequence can be made using this degeneracy to produce multiple nucleic acid sequences that all encode substantially the same protein.

[0111] The phrases "percent identity" and "% identity," as applied to polypeptide sequences, refer to the percentage of residue matches between at least two polypeptide sequences aligned using a standardized algorithm. Methods of polypeptide sequence alignment are well-known. Some alignment methods take into account conservative amino acid substitutions. Such conservative substitutions, explained in more detail above, generally preserve the charge and hydrophobicity at the site of substitution, thus preserving the structure (and therefore function) of the polypeptide.

[0112] Percent identity between polypeptide sequences may be determined using the default parameters of the CLUSTAL V algorithm as incorporated into the MEGALIGN version 3.12e sequence alignment program (described and referenced above). For pairwise alignments of polypeptide sequences using CLUSTAL V, the default parameters are set as follows: Ktuple=1, gap penalty=3, window=5, and "diagonals saved"=5. The PAM250 matrix is selected as the default residue weight table. As with polynucleotide alignments, the percent identity is reported by CLUSTAL V as the "percent similarity" between aligned polypeptide sequence pairs.

[0113] Alternatively the NCBI BLAST software suite may be used. For example, for a pairwise comparison of two polypeptide sequences, one may use the "BLAST 2 Sequences" tool Version 2.0.12 (Apr. 21, 2000) with blastp set at default parameters. Such default parameters may be, for example:

[0114] Matrix: BLOSUM62

[0115] Open Gap: 11 and Extension Gap: 1 penalties

[0116] Gap x drop-off. 50

[0117] Expect: 10

[0118] Word Size: 3

[0119] Filter: on

[0120] Percent identity may be measured over the length of an entire defined polypeptide sequence, for example, as defined by a particular SEQ ID number, or may be measured over a shorter length, for example, over the length of a fragment taken from a larger, defined polypeptide sequence, for instance, a fragment of at least 15, at least 20, at least 30, at least 40, at least 50, at least 70 or at least 150 contiguous residues. Such lengths are exemplary only, and it is understood that any fragment length supported by the sequences shown herein, in the tables, figures or Sequence Listing, may be used to describe a length over which percentage identity may be measured.

[0121] "Human artificial chromosomes" (HACs) are linear microchromosomes which may contain DNA sequences of about 6 kb to 10 Mb in size and which contain all of the elements required for chromosome replication, segregation and maintenance.

[0122] The term "humanized antibody" refers to an antibody molecule in which the amino acid sequence in the non-antigen binding regions has been altered so that the antibody more closely resembles a human antibody, and still retains its original binding ability.

[0123] "Hybridization" refers to the process by which a polynucleotide strand anneals with a complementary strand through base pairing under defined hybridization conditions. Specific hybridization is an indication that two nucleic acid sequences share a high degree of complementarity. Specific hybridization complexes form under permissive annealing conditions and remain hybridized after the "washing" step(s). The washing step(s) is particularly important in determining the stringency of the hybridization process, with more stringent conditions allowing less non-specific binding, i.e., binding between pairs of nucleic acid strands that are not perfectly matched. Permissive conditions for annealing of nucleic acid sequences are routinely determinable by one of ordinary skill in the art and may be consistent among hybridization experiments, whereas wash conditions may be varied among experiments to achieve the desired stringency, and therefore hybridization specificity. Permissive annealing conditions occur, for example, at 68.degree. C. in the presence of about 6.times.SSC, about 1% (w/v) SDS, and about 100 .mu.g/ml sheared, denatured salmon sperm DNA.

[0124] Generally, stringency of hybridization is expressed, in part, with reference to the temperature under which the wash step is carried out. Such wash temperatures are typically selected to be about 5.degree. C. to 20.degree. C. lower than the thermal melting point (T.sub.m) for the specific sequence at a defined ionic strength and pH. The T.sub.m is the temperature (under defined ionic strength and pH) at which 50% of the target sequence hybridizes to a perfectly matched probe. An equation for calculating T.sub.m and conditions for nucleic acid hybridization are well known and can be found in Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, 2.sup.nd ed., vol. 1-3, Cold Spring Harbor Press, Plainview N.Y.; specifically see volume 2, chapter 9.

[0125] High stringency conditions for hybridization between polynucleotides of the present invention include wash conditions of 68.degree. C. in the presence of about 0.2.times.SSC and about 0.1% SDS, for 1 hour. Alternatively, temperatures of about 65.degree. C., 60.degree. C., 55.degree. C., or 42.degree. C. may be used. SSC concentration may be varied from about 0.1 to 2.times.SSC, with SDS being present at about 0.1%. Typically, blocking reagents are used to block non-specific hybridization. Such blocking reagents include, for instance, sheared and denatured salmon sperm DNA at about 100-200 .mu.g/ml. Organic solvent, such as formamide at a concentration of about 35-50% v/v, may also be used under particular circumstances, such as for RNA:DNA hybridizations. Useful variations on these wash conditions will be readily apparent to those of ordinary skill in the art. Hybridization, particularly under high stringency conditions, may be suggestive of evolutionary similarity between the nucleotides. Such similarity is strongly indicative of a similar role for the nucleotides and their encoded polypeptides.

[0126] The term "hybridization complex" refers to a complex formed between two nucleic acid sequences by virtue of the formation of hydrogen bonds between complementary bases. A hybridization complex may be formed in solution (e.g., C.sub.0t or R.sub.0t analysis) or formed between one nucleic acid sequence present in solution and another nucleic acid sequence immobilized on a solid support (e.g., paper, membranes, filters, chips, pins or glass slides, or any other appropriate substrate to which cells or their nucleic acids have been fixed).

[0127] The words "insertion" and "addition" refer to changes in an amino acid or nucleotide sequence resulting in the addition of one or more amino acid residues or nucleotides, respectively.

[0128] "Immune response" can refer to conditions associated with inflammation, trauma, immune disorders, or infectious or genetic disease, etc. These conditions can be characterized by expression of various factors, e.g., cytokines, chemokines, and other signaling molecules, which may affect cellular and systemic defense systems.

[0129] An "immunogenic fragment" is a polypeptide or oligopeptide fragment of PKIN which is capable of eliciting an immune response when introduced into a living organism for example, a mammal. The term "immunogenic fragment" also includes any polypeptide or oligopeptide fragment of PKIN which is useful in any of the antibody production methods disclosed herein or known in the art.

[0130] The term "microarray" refers to an arrangement of a plurality of polynucleotides, polypeptides, or other chemical compounds on a substrate.

[0131] The terms "element" and "array element" refer to a polynucleotide, polypeptide, or other chemical compound having a unique and defined position on a microarray.

[0132] The term "modulate" refers to a change in the activity of PKIN. For example, modulation may cause an increase or a decrease in protein activity, binding characteristics, or any other biological, functional, or immunological properties of PKIN.

[0133] The phrases "nucleic acid" and "nucleic acid sequence" refer to a nucleotide, oligonucleotide, polynucleotide, or any fragment thereof. These phrases also refer to DNA or RNA of genomic or synthetic origin which may be single-stranded or double-stranded and may represent the sense or the antisense strand, to peptide nucleic acid (PNA), or to any DNA-like or RNA-like material.

[0134] "Operably linked" refers to the situation in which a first nucleic acid sequence is placed in a functional relationship with a second nucleic acid sequence. For instance, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Operably linked DNA sequences may be in close proximity or contiguous and, where necessary to join two protein coding regions, in the same reading frame.

[0135] "Peptide nucleic acid" (PNA) refers to an antisense molecule or anti-gene agent which comprises an oligonucleotide of at least about 5 nucleotides in length linked to a peptide backbone of amino acid residues ending in lysine. The terminal lysine confers solubility to the composition. PNAs preferentially bind complementary single stranded DNA or RNA and stop transcript elongation, and may be pegylated to extend their lifespan in the cell.

[0136] "Post-translational modification" of an PKIN may involve lipidation, glycosylation, phosphorylation, acetylation, racemization, proteolytic cleavage, and other modifications known in the art. These processes may occur synthetically or biochemically. Biochemical modifications will vary by cell type depending on the enzymatic milieu of PKIN.

[0137] "Probe" refers to nucleic acid sequences encoding PKIN, their complements, or fragments thereof, which are used to detect identical, allelic or related nucleic acid sequences. Probes are isolated oligonucleotides or polynucleotides attached to a detectable label or reporter molecule. Typical labels include radioactive isotopes, ligands, chemiluminescent agents, and enzymes. "Primers" are short nucleic acids, usually DNA oligonucleotides, which may be annealed to a target polynucleotide by complementary base-pairing. The primer may then be extended along the target DNA strand by a DNA polymerase enzyme. Primer pairs can be used for amplification (and identification) of a nucleic acid sequence, e.g., by the polymerase chain reaction (PCR).

[0138] Probes and primers as used in the present invention typically comprise at least 15 contiguous nucleotides of a known sequence. In order to enhance specificity, longer probes and primers may also be employed, such as probes and primers that comprise at least 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, or at least 150 consecutive nucleotides of the disclosed nucleic acid sequences. Probes and primers may be considerably longer than these examples, and it is understood that any length supported by the, specification, including the tables, figures, and Sequence Listing, may be used.

[0139] Methods for preparing and using probes and primers are described in the references, for example Sambrook, J. et al. (1989) Molecular Cloning: A Laboratory Manual, 2.sup.nd ed., vol. 1-3, Cold Spring Harbor Press, Plainview N.Y.; Ausubel, F. M. et al. (1987) Current Protocols in Molecular Biology, Greene Publ. Assoc. & Wiley-Intersciences, New York N.Y.; Innis, M. et al. (1990) PCR Protocols, A Guide to Methods and Applications, Academic Press, San Diego Calif. PCR primer pairs can be derived from a known sequence, for example, by using computer programs intended for that purpose such as Primer (Version 0.5, 1991, Whitehead Institute for Biomedical Research, Cambridge Mass.).

[0140] Oligonucleotides for use as primers are selected using software known in the art for such purpose. For example, OLIGO 4.06 software is useful for the selection of PCR primer pairs of up to 100 nucleotides each, and for the analysis of oligonucleotides and larger polynucleotides of up to 5,000 nucleotides from an input polynucleotide sequence of up to 32 kilobases. Similar primer selection programs have incorporated additional features for expanded capabilities. For example, the PrimOU primer selection program (available to the public from the Genome Center at University of Texas South West Medical Center, Dallas Tex.) is capable of choosing specific primers from megabase sequences and is thus useful for designing primers on a genome-wide scope. The Primer3 primer selection program (available to the public from the Whitehead Institute/MIT Center for Genome Research, Cambridge Mass.) allows the user to input a "mispriming library," in which sequences to avoid as primer binding sites are user-specified. Primer3 is useful, in particular, for the selection of oligonucleotides for microarrays. (The source code for the latter two primer selection programs may also be obtained from their respective sources and modified to meet the user's specific needs.) The PrimeGen program (available to the public from the UK Human Genome Mapping Project Resource Centre, Cambridge UTK) designs primers based on multiple sequence alignments, thereby allowing selection of primers that hybridize to either the most conserved or least conserved regions of aligned nucleic acid sequences. Hence, this program is useful for identification of both unique and conserved oligonucleotides and polynucleotide fragments. The oligonucleotides and polynucleotide fragments identified by any of the above selection methods are useful in hybridization technologies, for example, as PCR or sequencing primers, microarray elements, or specific probes to identify fully or partially complementary polynucleotides in a sample of nucleic acids. Methods of oligonucleotide selection are not limited to those described above.

[0141] A "recombinant nucleic acid" is a sequence that is not naturally occurring or has a sequence that is made by an artificial combination of two or more otherwise separated segments of sequence. This artificial combination is often accomplished by chemical synthesis or, more commonly, by the artificial manipulation of isolated segments of nucleic acids, e.g., by genetic engineering techniques such as those described in Sambrook, supra. The term recombinant includes nucleic acids that have been altered solely by addition, substitution, or deletion of a portion of the nucleic acid. Frequently, a recombinant nucleic acid may include a nucleic acid sequence operably linked to a promoter sequence. Such a recombinant nucleic acid may be part of a vector that is used, for example, to transform a cell.

[0142] Alternatively, such recombinant nucleic acids may be part of a viral vector, e.g., based on a vaccinia virus, that could be use to vaccinate a mammal wherein the recombinant nucleic acid is expressed, inducing a protective immunological response in the mammal.

[0143] A "regulatory element" refers to a nucleic acid sequence usually derived from untranslated regions of a gene and includes enhancers, promoters, introns, and 5' and 3' untranslated regions (UTRs). Regulatory elements interact with host or viral proteins which control transcription, translation, or RNA stability.

[0144] "Reporter molecules" are chemical or biochemical moieties used for labeling a nucleic acid, amino acid, or antibody. Reporter molecules include radionuclides; enzymes; fluorescent, chemiluminescent, or chromogenic agents; substrates; cofactors; inhibitors; magnetic particles; and other moieties known in the art.

[0145] An "RNA equivalent," in reference to a DNA sequence, is composed of the same linear sequence of nucleotides as the reference DNA sequence with the exception that all occurrences of the nitrogenous base thymine are replaced with uracil, and the sugar backbone is composed of ribose instead of deoxyribose.

[0146] The term "sample" is used in its broadest sense. A sample suspected of containing PKIN, nucleic acids encoding PKIN, or fragments thereof may comprise a bodily fluid; an extract from a cell, chromosome, organelle, or membrane isolated from a cell; a cell; genomic DNA, RNA, or cDNA, in solution or bound to a substrate; a tissue; a tissue print; etc.

[0147] The terms "specific binding" and "specifically binding" refer to that interaction between a protein or peptide and an agonist, an antibody, an antagonist, a small molecule, or any natural or synthetic binding composition. The interaction is dependent upon the presence of a particular structure of the protein, e.g., the antigenic determinant or epitope, recognized by the binding molecule. For example, if an antibody is specific for epitope "A," the presence of a polypeptide comprising the epitope A, or the presence of free unlabeled A, in a reaction containing free labeled A and the antibody will reduce the amount of labeled A that binds to the antibody.

[0148] The term "substantially purified" refers to nucleic acid or amino acid sequences that are removed from their natural environment and are isolated or separated, and are at least 60% free, preferably at least 75% free, and most preferably at least 90% free from other components with which they are naturally associated.

[0149] A "substitution" refers to the replacement of one or more amino acid residues or nucleotides by different amino acid residues or nucleotides, respectively.

[0150] "Substrate" refers to any suitable rigid or semi-rigid support including membranes, filters, chips, slides, wafers, fibers, magnetic or nonmagnetic beads, gels, tubing, plates, polymers, microparticles and capillaries. The substrate can have a variety of surface forms, such as wells, trenches, pins, channels and pores, to which polynucleotides or polypeptides are bound.

[0151] A "transcript image" refers to the collective pattern of gene expression by a particular cell type or tissue under given conditions at a given time.

[0152] "Transformation" describes a process by which exogenous DNA is introduced into a recipient cell. Transformation may occur under natural or artificial conditions according to various methods well known in the art, and may rely on any known method for the insertion of foreign nucleic acid sequences into a prokaryotic or eukaryotic host cell. The method for transformation is selected based on the type of host cell being transformed and may include, but is not limited to, bacteriophage or viral infection, electroporation, heat shock, lipofection, and particle bombardment. The term "transformed cells" includes stably transformed cells in which the inserted DNA is capable of replication either as an autonomously replicating plasmid or as part of the host chromosome, as well as transiently transformed cells which express the inserted DNA or RNA for limited periods of time.

[0153] A "transgenic organism," as used herein, is any organism, including but not limited to animals and plants, in which one or more of the cells of the organism contains heterologous nucleic acid introduced by way of human intervention, such as by transgenic techniques well known in the art. The nucleic acid is introduced into the cell, directly or indirectly by introduction into a precursor of the cell, by way of deliberate genetic manipulation, such as by microinjection or by infection with a recombinant virus. The term genetic manipulation does not include classical cross-breeding, or in vitro fertilization, but rather is directed to the introduction of a recombinant DNA molecule. The transgenic organisms contemplated in accordance with the present invention include bacteria, cyanobacteria, fungi, plants and animals. The isolated DNA of the present invention can be introduced into the host by methods known in the art, for example infection, transfection, transformation or transconjugation. Techniques for transferring the DNA of the present invention into such organisms are widely known and provided in references such as Sambrook et al. (1989), supra.

[0154] A "variant" of a particular nucleic acid sequence is defined as a nucleic acid sequence having at least 40% sequence identity to the particular nucleic acid sequence over a certain length of one of the nucleic acid sequences using blastn with the "BLAST 2 Sequences" tool Version 2.0.9 (May 7, 1999) set at default parameters. Such a pair of nucleic acids may show, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% or greater sequence identity over a certain defined length. A variant may be described as, for example, an "allelic" (as defined above), "splice," "species," or "polymorphic" variant. A splice variant may have significant identity to a reference molecule, but will generally have a greater or lesser number of polynucleotides due to alternative splicing of exons during mRNA processing. The corresponding polypeptide may possess additional functional domains or lack domains that are present in the reference molecule. Species variants are polynucleotide sequences that vary from one species to another. The resulting polypeptides will generally have significant amino acid identity relative to each other. A polymorphic variant is a variation in the polynucleotide sequence of a particular gene between individuals of a given species. Polymorphic variants also may encompass "single nucleotide polymorphisms" (SNPs) in which the polynucleotide sequence varies by one nucleotide base. The presence of SNPs may be indicative of, for example, a certain population, a disease state, or a propensity for a disease state.

[0155] A "variant" of a particular polypeptide sequence is defined as a polypeptide sequence having at least 40% sequence identity to the particular polypeptide sequence over a certain length of one of the polypeptide sequences using blastp with the "BLAST 2 Sequences" tool Version 2.0.9 (May 7, 1999) set at default parameters. Such a pair of polypeptides may show, for example, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% or greater sequence identity over a certain defined length of one of the polypeptides.

[0156] The Invention

[0157] The invention is based on the discovery of new human kinases (PKIN), the polynucleotides encoding PKIN, and the use of these compositions for the diagnosis, treatment, or prevention of cancer, immune disorders, disorders affecting growth and development, cardiovascular diseases, and lipid disorders.

[0158] Table 1 summarizes the nomenclature for the full length polynucleotide and polypeptide sequences of the invention. Each polynucleotide and its corresponding polypeptide are correlated to a single Incyte project identification number (Incyte Project ID). Each polypeptide sequence is denoted by both a polypeptide sequence identification number (Polypeptide SEQ ID NO:) and an Incyte polypeptide sequence number (Incyte Polypeptide ID) as shown. Each polynucleotide sequence is denoted by both a polynucleotide sequence identification number (Polynucleotide SEQ ID NO:) and an Incyte polynucleotide consensus sequence number (Incyte Polynucleotide ID) as shown.

[0159] Table 2 shows sequences with homology to the polypeptides of the invention as identified by BLAST analysis against the GenBank protein (genpept) database. Columns 1 and 2 show the polypeptide sequence identification number (Polypeptide SEQ ID NO:) and the corresponding Incyte polypeptide sequence number (Incyte Polypeptide ID) for polypeptides of the invention. Column 3 shows the GenBank identification number (Genbank ID NO:) of the nearest GenBank homolog. Column 4 shows the probability score for the match between each polypeptide and its GenBank homolog. Column S shows the annotation of the GenBank homolog along with relevant citations where applicable, all of which are expressly incorporated by reference herein.

[0160] Table 3 shows various structural features of the polypeptides of the invention. Columns 1 and 2 show the polypeptide sequence identification number (SEQ ID NO:) and the corresponding Incyte polypeptide sequence number (Incyte Polypeptide ID) for each polypeptide of the invention. Column 3 shows the number of amino acid residues in each polypeptide. Column 4 shows potential phosphorylation sites, and column 5 shows potential glycosylation sites, as determined by the MOTIFS program of the GCG sequence analysis software package (Genetics Computer Group, Madison Wis.). Column 6 shows amino acid residues comprising signature sequences, domains, and motifs. Column 7 shows analytical methods for protein structure/function analysis and in some cases, searchable databases to which the analytical methods were applied.

[0161] Together, Tables 2 and 3 summarize the properties of polypeptides of the invention, and these properties establish that the claimed polypeptides are human kinases. For example, SEQ ID NO:12 is 95% identical to a human adenylate kinase (GenBank ID g28577) as determined by the Basic Local Alignment Search Tool (BLAST). (See Table 2). The BLAST probability score is 2.4e-112, which indicates the probability of obtaining the observed polypeptide sequence alignment by chance. Data from BLIMPS, MOTIFS, and PROFILE analysis; from BLAST analysis using the DOMO and PRODOM databases; and from HMMER analysis using the PFAM database further support the categorization of SEQ ID NO:12 as an adenylate kinase. (See Table 3). In an alternative example, SEQ ID NO:13 is 92% identical to rat PCTAIRE 3 (GenBank ID g2257588) as determined by the Basic Local Alignment Search Tool (BLAST). The BLAST probability score is 1.4e-210, which indicates the probability of obtaining the observed polypeptide sequence alignment by chance (Table 2). PCTAIRE 1, 2, and 3 comprise a subfamily of Cdc2-related kinases that are primarily expressed in post-mitotic cells. PCTAIRE 2 and PCTAIRE 3 are expressed in the brain (Hirose, T. et al., (1997) Eur. J. Biochem. 249:481-488; Okada, T. et al., (1992) Oncogene 7:2249-2258). SEQ ID NO:13 also contains a kinase active site domain as determined by searching for statistically significant matches in the hidden Markov model (HMM)-based PFAM database of conserved protein family domains. (See Table 3.) Data from BLIMPS, MOTIFS, and PROPILESCAN analyses provide further corroborative evidence that SEQ ID NO:13 is a kinase. In another alternative example, SEQ ID NO:18 is 86% identical to human cell cycle related kinase (GenBank ID g4090958) as determined by the Basic Local Alignment Search Tool (BLAST). (See Table 2.) The BLAST probability score is 9.2e-85, which indicates the probability of obtaining the observed polypeptide sequence alignment by chance. SEQ ID NO:18 also contains a eukaryotic protein kinase domain as determined by searching for statistically significant matches in the hidden Markov model (HMM)-based PFAM database of conserved protein family domains. (See Table 3.) Data from BLIMPS, MOTIFS, and PROFILESCAN analyses provide further corroborative evidence that SEQ ID NO:18 is a cell cycle related kinase. SEQ ID NO:1-11 and SEQ ID NO:14-17 were analyzed and annotated in a similar manner. The algorithms and parameters for the analysis of SEQ ID NO:1-18 are described in Table 7.

[0162] As shown in Table 4, the full length polynucleotide sequences of the present invention were assembled using cDNA sequences or coding (exon) sequences derived from genomic DNA, or any combination of these two types of sequences. Columns 1 and 2 list the polynucleotide sequence identification number (Polynucleotide SEQ ID NO:) and the corresponding Incyte polynucleotide consensus sequence number (Incyte Polynucleotide ID) for each polynucleotide of the invention. Column 3 shows the length of each polynucleotide sequence in basepairs. Column 4 lists fragments of the polynucleotide sequences which are useful, for example, in hybridization or amplification technologies that identify SEQ ID NO:19-36 or that distinguish between SEQ ID NO:19-36 and related polynucleotide sequences. Column 5 shows identification numbers corresponding to cDNA sequences, coding sequences (exons) predicted from genomic DNA, and/or sequence assemblages comprised of both cDNA and genomic DNA. These sequences were used to assemble the full length polynucleotide sequences of the invention. Columns 6 and 7 of Table 4 show the nucleotide start (5') and stop (3') positions of the cDNA and genomic sequences in column 5 relative to their respective full length sequences.

[0163] The identification numbers in Column 5 of Table 4 may refer specifically, for example, to Incyte cDNAs along with their corresponding cDNA libraries. For example, 6311370H1 is the identification number of an Incyte cDNA sequence, and NERDTDN03 is the cDNA library from which it is derived. Incyte cDNAs for which cDNA libraries are not indicated were derived from pooled cDNA libraries (e.g., 70518523D1). Alternatively, the identification numbers in 5 may refer to GenBank cDNAs or ESTs (e.g., g1860144) which contributed to the assembly of the full length polynucleotide sequences. Alternatively, the identification numbers in column 5 may refer to coding regions predicted by Genscan analysis of genomic DNA. For example, GNN.g5924006.sub.--004.edit is the identification number of a Genscan-predicted coding sequence, with g5924006 being the GenBank identification number of the sequence to which Genscan was applied. The Genscan-predicted coding sequences may have been edited prior to assembly. (See Example IV.) Alternatively, the identification numbers in column 5 may refer to assemblages of both cDNA and Genscan-predicted exons brought together by an "exon stitching" algorithm. (See Example V.) Alternatively, the identification numbers in column 5 may refer to assemblages of both cDNA and Genscan-predicted exons brought together by an "exon-stretching" algorithm. (See Example V.) In some cases, Incyte cDNA coverage redundant with the sequence coverage shown in column 5 was obtained to confirm the final consensus polynucleotide sequence, but the relevant Incyte cDNA identification numbers are not shown.

[0164] Table 5 shows the representative cDNA libraries for those full length polynucleotide sequences which were assembled using Incyte cDNA sequences. The representative cDNA library is the Incyte cDNA library which is most frequently represented by the Incyte cDNA sequences which were used to assemble and confirm the above polynucleotide sequences. The tissues and vectors which were used to construct the cDNA libraries shown in Table 5 are described in Table 6.

[0165] The invention also encompasses PKIN variants. A preferred PKIN variant is one which has at least about 80%, or alternatively at least about 96%, or alternatively at least about 95%, or even at least about 98% amino acid sequence identity to the PKIN amino acid sequence, and which contains at least one functional or structural characteristic of PKIN.

[0166] The invention also encompasses polynucleotides which encode PKIN. In a particular embodiment, the invention encompasses a polynucleotide sequence comprising a sequence selected from the group consisting of SEQ ID NO:19-36, which encodes PKIN. The polynucleotide sequences of SEQ ID NO:19-36, as presented in the Sequence Listing, embrace the equivalent RNA sequences, wherein occurrences of the nitrogenous base thymine are replaced with uracil, and the sugar backbone is composed of ribose instead of deoxyribose.

[0167] The invention also encompasses a variant of a polynucleotide sequence encoding PKIN. In particular, such a variant polynucleotide sequence will have at least about 70%, or alternatively at least about 85%, or alternatively at least about 95%, or even at least about 98% polynucleotide sequence identity to the polynucleotide sequence encoding PKIN. A particular aspect of the invention encompasses a variant of a polynucleotide sequence comprising a sequence selected from the group consisting of SEQ ID NO:19-36 which has at least about 70%, or alternatively at least about 85%, or alternatively at least about 95%, or even at least about 98% polynucleotide sequence identity to a nucleic acid sequence selected from the group consisting of SEQ ID NO:19-36. Any one of the polynucleotide variants described above can encode an amino acid sequence which contains at least one functional or structural characteristic of PKIN.

[0168] It will be appreciated by those skilled in the art that as a result of the degeneracy of the genetic code, a multitude of polynucleotide sequences encoding PKIN, some bearing minimal similarity to the polynucleotide sequences of any known and naturally occurring gene, may be produced. Thus, the invention contemplates each and every possible variation of polynucleotide sequence that could be made by selecting combinations based on possible codon choices. These combinations are made in accordance with the standard triplet genetic code as applied to the polynucleotide sequence of naturally occurring PKIN, and all such variations are to be considered as being specifically disclosed.

[0169] Although nucleotide sequences which encode PKIN and its variants are generally capable of hybridizing to the nucleotide sequence of the naturally occurring PKIN under appropriately selected conditions of stringency, it may be advantageous to produce nucleotide sequences encoding PKIN or its derivatives possessing a substantially different codon usage, e.g., inclusion of non-naturally occurring codons. Codons may be selected to increase the rate at which expression of the peptide occurs in a particular prokaryotic or eukaryotic host in accordance with the frequency with which particular codons are utilized by the host. Other reasons for substantially altering the nucleotide sequence encoding PEIN and its derivatives without altering the encoded amino acid sequences include the production of RNA transcripts having more desirable properties, such as a greater half-life, than transcripts produced from the naturally occurring sequence.

[0170] The invention also encompasses production of DNA sequences which encode PKIN and PEIN derivatives, or fragments thereof, entirely by synthetic chemistry. After production, the synthetic sequence may be inserted into any of the many available expression vectors and cell systems using reagents well known in the art. Moreover, synthetic chemistry may be used to introduce mutations into a sequence encoding PKIN or any fragment thereof.

[0171] Also encompassed by the invention are polynucleotide sequences that are capable of hybridizing to the claimed polynucleotide sequences, and, in particular, to those shown in SEQ ID NO:19-36 and fragments thereof under various conditions of stringency. (See, e.g., Wahl, G. M. and S. L. Berger (1987) Methods Enzymol. 152:399407; Kimmel, A. R. (1987) Methods Enzymol. 152:507-511.) Hybridization conditions, including annealing and wash conditions, are described in "Definitions."

[0172] Methods for DNA sequencing are well known in the art and may be used to practice any of the embodiments of the invention. The methods may employ such enzymes as the Klenow fragment of DNA polymerase 1, SEQUENASE (US Biochemical, Cleveland Ohio), Taq polymerase (Applied Biosystems), thermostable T7 polymerase (Amersham Pharmacia Biotech, Piscataway N.J.), or combinations of polymerases and proofreading exonucleases such as those found in the ELONGASE amplification system (Life Technologies, Gaithersburg Md.). Preferably, sequence preparation is automated with machines such as the MJCROLAB 2200 liquid transfer system (Hamilton, Reno Nev.), PTC200 thermal cycler (MJ Research, Watertown Mass.) and ABI CATALYST 800 thermal cycler (Applied Biosystems). Sequencing is then carried out using either the ABI 373 or 377 DNA sequencing system (Applied Biosystems), the MEGABACE 1000 DNA sequencing system (Molecular Dynamics, Sunnyvale Calif.), or other systems known in the art. The resulting sequences are analyzed using a variety of algorithms which are well known in the art. (See, e.g., Ausubel, F. M. (1997) Short Protocols in Molecular Biology, John Wiley & Sons, New York N.Y., unit 7.7; Meyers, R. A. (1995) Molecular Biology and Biotechnology, Wiley VCH, New York N.Y., pp. 856-853.)

[0173] The nucleic acid sequences encoding PKIN may be extended utilizing a partial nucleotide sequence and employing various PCR-based methods known in the art to detect upstream sequences, such as promoters and regulatory elements. For example, one method which may be employed, restriction-site PCR, uses universal and nested primers to amplify unknown sequence from genomic DNA within a cloning vector. (See, e.g., Sarkar, G. (1993) PCR Methods Applic. 2:318-322.) Another method, inverse PCR, uses primers that extend in divergent directions to amplify unknown sequence from a circularized template. The template is derived from restriction fragments comprising a known genomic locus and surrounding sequences. (See, e.g., Triglia, T. et al. (1988) Nucleic Acids Res. 16:8186.) A third method, capture PCR, involves PCR amplification of DNA fragments adjacent to known sequences in human and yeast artificial chromosome DNA. (See, e.g., Lagerstrom, M. et al. (1991) PCR Methods Applic. 1:111-119.) In this method, multiple restriction enzyme digestions and ligations may be used to insert an engineered double-stranded sequence into a region of unknown sequence before performing PCR. Other methods which may be used to retrieve unknown sequences are known in the art. (See, e.g., Parker, J. D. et al. (1991) Nucleic Acids Res. 19:3055-3060). Additionally, one may use PCR, nested primers, and PROMOTERFINDER libraries (Clontech, Palo Alto Calif.) to walk genomic DNA. This procedure avoids the need to screen libraries and is useful in finding intron/exon junctions. For all PCR-based methods, primers may be designed using commercially available software, such as OLIGO 4.06 primer analysis software (National Biosciences, Plymouth Minn.) or another appropriate program, to be about 22 to 30 nucleotides in length, to have a GC content of about 50% or more, and to anneal to the template at temperatures of about 68.degree. C. to 72.degree. C.

[0174] When screening for full length cDNAs, it is preferable to use libraries that have been size-selected to include larger cDNAs. In addition, random-primed libraries, which often include sequences containing the 5' regions of genes, are preferable for situations in which an oligo d(T) library does not yield a full-length cDNA. Genomic libraries may be useful for extension of sequence into 5' non-transcribed regulatory regions.

[0175] Capillary electrophoresis systems which are commercially available may be used to analyze the size or confirm the nucleotide sequence of sequencing or PCR products. In particular, capillary sequencing may employ flowable polymers for electrophoretic separation, four different nucleotide-specific, laser-stimulated fluorescent dyes, and a charge coupled device camera for detection of the emitted wavelengths. Output/light intensity may be converted to electrical signal using appropriate software (e.g., GENOTYPER and SEQUENCE NAVIGATOR, Applied Biosystems), and the entire process from loading of samples to computer analysis and electronic data display may be computer controlled. Capillary electrophoresis is especially preferable for sequencing small DNA fragments which may be present in limited amounts in a particular sample.

[0176] In another embodiment of the invention, polynucleotide sequences or fragments thereof which encode PKIN may be cloned in recombinant DNA molecules that direct expression of PKIN, or fragments or functional equivalents thereof, in appropriate host cells. Due to the inherent degeneracy of the genetic code, other DNA sequences which encode substantially the same or a functionally equivalent amino acid sequence may be produced and used to express PKIN.

[0177] The nucleotide sequences of the present invention can be engineered using methods generally known in the art in order to alter PKIN-encoding sequences for a variety of purposes including, but not limited to, modification of the cloning, processing, and/or expression of the gene product. DNA shuffling by random fragmentation and PCR reassembly of gene fragments and synthetic oligonucleotides may be used to engineer the nucleotide sequences. For example, oligonucleotide-mediated site-directed mutagenesis may be used to introduce mutations that create new restriction sites, alter glycosylation patterns, change codon preference, produce splice variants, and so forth.

[0178] The nucleotides of the present invention may be subjected to DNA shuffling techniques such as MOLECULARBREEDING (Maxygen Inc., Santa Clara Calif.; described in U.S. Pat. No. 5,837,458; Chang, C.sub.1-C. et al. (1999) Nat. Biotechnol. 17:793-797; Christians, F. C. et al. (1999) Nat. Biotechnol. 17:259-264; and Crameri, A. et al. (1996) Nat. Biotechnol. 14:315-319) to alter or improve the biological properties of PKIN, such as its biological or enzymatic activity or its ability to bind to other molecules or compounds. DNA shuffling is a process by which a library of gene variants is produced using PCR-mediated recombination of gene fragments. The library is then subjected to selection or screening procedures that identify those gene variants with the desired properties. These preferred variants may then be pooled and further subjected to recursive rounds of DNA shuffling and selection/screening. Thus, genetic diversity is created through "artificial" breeding and rapid molecular evolution. For example, fragments of a single gene containing random point mutations may be recombined, screened, and then reshuffled until the desired properties are optimized. Alternatively, fragments of a given gene may be recombined with fragments of homologous genes in the same gene family, either from the same or different species, thereby maximizing the genetic diversity of multiple naturally occurring genes in a directed and controllable manner.

[0179] In another embodiment, sequences encoding PKIN may be synthesized, in whole or in part, using chemical methods well known in the art. (See, e.g., Caruthers, M. H. et al. (1980) Nucleic Acids Symp. Ser. 7:215-223; and Horn, T. et al. (1980) Nucleic Acids Symp. Ser. 7:225-232.) Alternatively, PKIN itself or a fragment thereof may be synthesized using chemical methods. For example, peptide synthesis can be performed using various solution-phase or solid-phase techniques. (See, e.g., Creighton, T. (1984) Proteins, Structures and Molecular Properties, W H Freeman, New York N.Y., pp. 55-60; and Roberge, J. Y. et al. (1995) Science 269:202-204.) Automated synthesis may be achieved using the ABI 431A peptide synthesizer (Applied Biosystems). Additionally, the amino acid sequence of PKIN, or any part thereof, may be altered during direct synthesis and/or combined with sequences from other proteins, or any part thereof, to produce a variant polypeptide or a polypeptide having a sequence of a naturally occurring polypeptide.

[0180] The peptide may be substantially purified by preparative high performance liquid chromatography. (See, e.g., Chiez, R. M. and F. Z. Regnier (1990) Methods Enzymol. 182:392-421.) The composition of the synthetic peptides may be confirmed by amino acid analysis or by sequencing. (See, e.g., Creighton, supra, pp. 28-53.)

[0181] In order to express a biologically active PKIN, the nucleotide sequences encoding PKIN or derivatives thereof may be inserted into an appropriate expression vector, i.e., a vector which contains the necessary elements for transcriptional and translational control of the inserted coding sequence in a suitable host. These elements include regulatory sequences, such as enhancers, constitutive and inducible promoters, and 5' and 3' untranslated regions in the vector and in polynucleotide sequences encoding PKIN. Such elements may vary in their strength and specificity. Specific initiation signals may also be used to achieve more efficient translation of sequences encoding PKIN. Such signals include the ATG initiation codon and adjacent sequences, e.g. the Kozak sequence. In cases where sequences encoding PKIN and its initiation codon and upstream regulatory sequences are inserted into the appropriate expression vector, no additional transcriptional or translational control signals may be needed. However, in cases where only coding sequence, or a fragment thereof, is inserted, exogenous translational control signals including an in-frame ATG initiation codon should be provided by the vector. Exogenous translational elements and initiation codons may be of various origins, both natural and synthetic. The efficiency of expression may be enhanced by the inclusion of enhancers appropriate for the particular host cell system used. (See, e.g., Scharf, D. et al. (1994) Results Probl. Cell Differ. 20:125-162.)

[0182] Methods which are well known to those skilled in the art may be used to construct expression vectors containing sequences encoding PKIN and appropriate transcriptional and translational control elements. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. (See, e.g., Sambrook, J. et al. (1989) Molecular Cloning A Laboratory Manual, Cold Spring Harbor Press, Plainview N.Y., ch. 4, 8, and 16-17; Ausubel, F. M. et al. (1995) Current Protocols in Molecular Biology, John Wiley & Sons, New York N.Y., ch. 9, 13, and 16.)

[0183] A variety of expression vector/host systems may be utilized to contain and express sequences encoding PKIN. These include, but are not limited to, microorganisms such as bacteria transformed with recombinant bacteriophage, plasmid, or cosmid DNA expression vectors; yeast transformed with yeast expression vectors; insect cell systems infected with viral expression vectors (e.g., baculovirus); plant cell systems transformed with viral expression vectors (e.g., cauliflower mosaic virus, CaMV, or tobacco mosaic virus, TMV) or with bacterial expression vectors (e.g., Ti or pBR322 plasmids); or animal cell systems. (See, e.g., Sambrook, supra; Ausubel, supra; Van Heeke, G. and S. M. Schuster (1989) J. Biol. Chem. 264:5503-5509; Engelhard, E. K. et al. (1994) Proc. Natl. Acad. Sci. USA 91:3224-3227; Sandig, V. et al. (1996) Hum. Gene Ther. 7:1937-1945; Takamatsu, N. (1987) EMBO J. 6:307-311; The McGraw Hill Yearbook of Science and Technology (1992) McGraw Hill, New York N.Y., pp. 191-196; Logan, J. and T. Shenk (1984) Proc. Natl. Acad. Sci. USA 81:3655-3659; and Harrington, J. J. et al. (1997) Nat. Genet. 15:345-355.) Expression vectors derived from retroviruses, adenoviruses, or herpes or vaccinia viruses, or from various bacterial plasmids, may be used for delivery of nucleotide sequences to the targeted organ, tissue, or cell population. (See, e.g., Di Nicola, M. et al. (1998) Cancer Gen. Ther. 5(6):350-356; Yu, M. et al. (1993) Proc. Natl. Acad. Sci. USA 90(13):6340-6344; Buller, R. M. et al. (1985) Nature 317(6040):813-815; McGregor, D. P. et al. (1994) Mol. Immunol. 31(3):219-226; and Verma, I. M. and N. Somia (1997) Nature 389:239-242.) The invention is not limited by the host cell employed.

[0184] In bacterial systems, a number of cloning and expression vectors may be selected depending upon the use intended for polynucleotide sequences encoding PKIN. For example, routine cloning, subcloning, and propagation of polynucleotide sequences encoding PKIN can be achieved using a multifunctional E. coli vector such as PBLUESCRIPT (Stratagene, La Jolla Calif.) or PSPORT1 plasmid (Life Technologies). Ligation of sequences encoding PKIN into the vector's multiple cloning site disrupts the lacZ gene, allowing a colorimetric screening procedure for identification of transformed bacteria containing recombinant molecules. In addition, these vectors may be useful for in vitro transcription, dideoxy sequencing, single strand rescue with helper phage, and creation of nested deletions in the cloned sequence. (See, e.g., Van Heeke, G. and S. M. Schuster (1989) J. Biol. Chem. 264:5503-5509.) When large quantities of PKIN are needed, e.g. for the production of antibodies, vectors which direct high level expression of PKIN may be used. For example, vectors containing the strong, inducible SP6 or T7 bacteriophage promoter may be used.

[0185] Yeast expression systems may be used for production of PKIN. A number of vectors containing constitutive or inducible promoters, such as alpha factor, alcohol oxidase, and PGH promoters, may be used in the yeast Saccharomyces cerevisiae or Pichia pastoris. In addition, such vectors direct either the secretion or intracellular retention of expressed proteins and enable integration of foreign sequences into the host genome for stable propagation. (See, e.g., Ausubel, 1995, supra; Bitter, G. A. et al. (1987) Methods Enzymol. 153:516-544; and Scorer, C. A. et al. (1994) Bio/Technology 12:181-184.)

[0186] Plant systems may also be used for expression of PKIN. Transcription of sequences encoding PKIN may be driven by viral promoters, e.g., the 35S and 19S promoters of CaMV used alone or in combination with the omega leader sequence from TMV (Takamatsu, N. (1987) EMBO J. 6:307-311). Alternatively, plant promoters such as the small subunit of RUBISCO or heat shock promoters may be used. (See, e.g., Coruzzi, G. et al. (1984) EMBO J. 3:1671-1680; Broglie, R. et al. (1984) Science 224:838-843; and Winter, J. et al. (1991) Results Probl. Cell Differ. 17:85-105.) These constructs can be introduced into plant cells by direct DNA transformation or pathogen-mediated transfection. (See, e.g., The McGraw Hill Yearbook of Science and Technology (1992) McGraw Hill, New York N.Y., pp. 191-196.)

[0187] In mammalian cells, a number of viral-based expression systems may be utilized. In cases where an adenovirus is used as an expression vector, sequences encoding PKIN may be ligated into an adenovirus transcription/translation complex consisting of the late promoter and tripartite leader sequence. Insertion in a non-essential E1 or E3 region of the viral genome may be used to obtain infective virus which expresses PKIN in host cells. (See, e.g., Logan, J. and T. Shenk (1984) Proc. Natl. Acad. Sci. USA 81:3655-3659.) In addition, transcription enhancers, such as the Rous sarcoma virus (RSV) enhancer, may be used to increase expression in mammalian host cells. SV40 or EBV-based vectors may also be used for high-level protein expression.

[0188] Human artificial chromosomes (HACs) may also be employed to deliver larger fragments of DNA than can be contained in and expressed from a plasmid. HACs of about 6 kb to 10 Mb are constructed and delivered via conventional delivery methods (liposomes, polycationic amino polymers, or vesicles) for therapeutic purposes. (See, e.g., Harrington, J. J. et al. (1997) Nat. Genet. 15:345-355.)

[0189] For long term production of recombinant proteins in mammalian systems, stable expression of PKIN in cell lines is preferred. For example, sequences encoding PKIN can be transformed into cell lines using expression vectors which may contain viral origins of replication and/or endogenous expression elements and a selectable marker gene on the same or on a separate vector. Following the introduction of the vector, cells may be allowed to grow for about 1 to 2 days in enriched media before being switched to selective media. The purpose of the selectable marker is to confer resistance to a selective agent, and its presence allows growth and recovery of cells which successfully express the introduced sequences. Resistant clones of stably transformed cells may be propagated using tissue culture techniques appropriate to the cell type.

[0190] Any number of selection systems may be used to recover transformed cell lines. These include, but are not limited to, the herpes simplex virus thymidine kinase and adenine phosphoribosyltransferase genes, for use in tk.sup.- and apr.sup.- cells, respectively. (See, e.g., Wigler, M. et al. (1977) Cell 11:223-232; Lowy, I. et al. (1980) Cell 22:817-823.) Also, antimetabolite, antibiotic, or herbicide resistance can be used as the basis for selection. For example, dhfr confers resistance to methotrexate; neo confers resistance to the aminoglycosides neomycin and G418; and als and pat confer resistance to chlorsulfuron and phosphinotricin acetyltransferase, respectively. (See, e.g., Wigler, M. et al. (1980) Proc. Natl. Acad. Sci. USA 77:3567-3570; Colbere-Garapin, F. et al. (1981) J. Mol. Biol. 150:1-14.) Additional selectable genes have been described, e.g., trpB and hisD, which alter cellular requirements for metabolites. (See, e.g., Hartman, S. C. and R. C. Mulligan (1988) Proc. Natl. Acad. Sci. USA 85:8047-8051.) Visible markers, e.g., anthocyanins, green fluorescent proteins (GFP; Clontech), B glucuronidase and its substrate B-glucuronide, or luciferase and its substrate luciferin may be used. These markers can be used not only to identify transformants, but also to quantify the amount of transient or stable protein expression attributable to a specific vector system. (See, e.g., Rhodes, C. A. (1995) Methods Mol. Biol. 55:121-131.)

[0191] Although the presence/absence of marker gene expression suggests that the gene of interest is also present, the presence and expression of the gene may need to be confirmed. For example, if the sequence encoding PEIN is inserted within a marker gene sequence, transformed cells containing sequences encoding PKIN can be identified by the absence of marker gene function. Alternatively, a marker gene can be placed in tandem with a sequence encoding PKIN under the control of a single promoter. Expression of the marker gene in response to induction or selection usually indicates expression of the tandem gene as well.

[0192] In general, host cells that contain the nucleic acid sequence encoding PKIN and that express PKIN may be identified by a variety of procedures known to those of skill in the art. These procedures include, but are not limited to, DNA-DNA or DNA-RNA hybridizations, PCR amplification, and protein bioassay or immunoassay techniques which include membrane, solution, or chip based technologies for the detection and/or quantification of nucleic acid or protein sequences.

[0193] Immunological methods for detecting and measuring the expression of PKIN using either specific polyclonal or monoclonal antibodies are known in the art. Examples of such techniques include enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs), and fluorescence activated cell sorting (FACS). A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering epitopes on PKIN is preferred, but a competitive binding assay may be employed. These and other assays are well known in the art. (See, e.g., Hampton, R. et al. (1990) Serological Methods, a Laboratory Manual, APS Press, St. Paul Minn., Sect. IV; Coligan, J. E. et al. (1997) Current Protocols in Immunology, Greene Pub. Associates and Wiley-Interscience, New York N.Y.; and Pound, J. D. (1998) Immunochemical Protocols, Humana Press, Totowa N.J.)

[0194] A wide variety of labels and conjugation techniques are known by those skilled in the art and may be used in various nucleic acid and amino acid assays. Means for producing labeled hybridization or PCR probes for detecting sequences related to polynucleotides encoding PKIN include oligolabeling, nick translation, end-labeling, or PCR amplification using a labeled nucleotide. Alternatively, the sequences encoding PKIN, or any fragments thereof, may be cloned into a vector for the production of an mRNA probe. Such vectors are known in the art, are commercially available, and may be used to synthesize RNA probes in vitro by addition of an appropriate RNA polymerase such as T7, T3, or SP6 and labeled nucleotides. These procedures may be conducted using a variety of commercially available kits, such as those provided by Amersham Pharmacia Biotech, Promega (Madison Wis.), and US Biochemical. Suitable reporter molecules or labels which may be used for ease of detection include radionuclides, enzymes, fluorescent, chemiluminescent, or chromogenic agents, as well as substrates, cofactors, inhibitors, magnetic particles, and the like.

[0195] Host cells transformed with nucleotide sequences encoding PKIN may be cultured under conditions suitable for the expression and recovery of the protein from cell culture. The protein produced by a transformed cell may be secreted or retained intracellularly depending on the sequence and/or the vector used. As will be understood by those of skill in the art, expression vectors containing polynucleotides which encode PKIN may be designed to contain signal sequences which direct secretion of PKIN through a prokaryotic or eukaryotic cell membrane.

[0196] In addition, a host cell strain may be chosen for its ability to modulate expression of the inserted sequences or to process the expressed protein in the desired fashion. Such modifications of the polypeptide include, but are not limited to, acetylation, carboxylation, glycosylation, phosphorylation, lipidation, and acylation. Post-translational processing which cleaves a "prepro" or "pro" form of the protein may also be used to specify protein targeting, folding, and/or activity. Different host cells which have specific cellular machinery and characteristic mechanisms for post-translational activities (e.g., CHO, HeLa, MDCK, HBEK293, and WI38) are available from the American Type Culture Collection (ATCC, Manassas Va.) and may be chosen to ensure the correct modification and processing of the foreign protein.

[0197] In another embodiment of the invention, natural, modified, or recombinant nucleic acid sequences encoding PKIN may be ligated to a heterologous sequence resulting in translation of a fusion protein in any of the aforementioned host systems. For example, a chimeric PKIN protein containing a heterologous moiety that can be recognized by a commercially available antibody may facilitate the screening of peptide libraries for inhibitors of PKIN activity. Heterologous protein and peptide moieties may also facilitate purification of fusion proteins using commercially available affinity matrices. Such moieties include, but are not limited to, glutathione S-transferase (GST), maltose binding protein (MBP), thioredoxin (Trx), calmodulin binding peptide (CBP), 6-His, FLAG, c-myc, and hemagglutinin (HA). GST, MBP, Trx, CBP, and 6-His enable purification of their cognate fusion proteins on immobilized glutathione, maltose, phenylarsine oxide, calmodulin, and metal-chelate resins, respectively. FLAG, c-myc, and hemagglutinin (HA) enable immunoaffinity purification of fusion proteins using commercially available monoclonal and polyclonal antibodies that specifically recognize these epitope tags. A fusion protein may also be engineered to contain a proteolytic cleavage site located between the PKIN encoding sequence and the heterologous protein sequence, so that PKIN may be cleaved away from the heterologous moiety following purification. Methods for fusion protein expression and purification are discussed in Ausubel (1995, supra, ch. 10). A variety of commercially available kits may also be used to facilitate expression and purification of fusion proteins.

[0198] In a further embodiment of the invention, synthesis of radiolabeled PKIN may be achieved in: vitro using the TNT rabbit reticulocyte lysate or wheat germ extract system (Promega). These systems couple transcription and translation of protein-coding sequences operably associated with the T7, T3, or SP6 promoters. Translation takes place in the presence of a radiolabeled amino acid precursor, for example, .sup.35S-methionine.

[0199] PKIN of the present invention or fragments thereof may be used to screen for compounds that specifically bind to PKIN. At least one and up to a plurality of test compounds may be screened for specific binding to PKIN. Examples of test compounds include antibodies, oligonucleotides, proteins (e.g., receptors), or small molecules.

[0200] In one embodiment, the compound thus identified is closely related to the natural ligand of PKIN, e.g., a ligand or fragment thereof, a natural substrate, a structural or functional mimetic, or a natural binding partner. (See, e.g., Coligan, J. E. et al. (1991) Current Protocols in Immunology 1(2): Chapter 5.) Similarly, the compound can be closely related to the natural receptor to which PKIN binds, or to at least a fragment of the receptor, e.g., the ligand binding site. In either case, the compound can be rationally designed using known techniques. In one embodiment, screening for these compounds involves producing appropriate cells which express PKIN, either as a secreted protein or on the cell membrane. Preferred cells include cells from mammals, yeast, Drosophila, or E. coli. Cells expressing PEIN or cell membrane fractions which contain PKIN are then contacted with a test compound and binding, stimulation, or inhibition of activity of either PKIN or the compound is analyzed.

[0201] An assay may simply test binding of a test compound to the polypeptide, wherein binding is detected by a fluorophore, radioisotope, enzyme conjugate, or other detectable label. For example, the assay may comprise the steps of combining at least one test compound with PKIN, either in solution or affixed to a solid support, and detecting the binding of PKIN to the compound. Alternatively, the assay may detect or measure binding of a test compound in the presence of a labeled competitor. Additionally, the assay may be carried out using cell-free preparations, chemical libraries, or natural product mixtures, and the test compound(s) may be free in solution or affixed to a solid support.

[0202] PKIN of the present invention or fragments thereof may be used to screen for compounds that modulate the activity of PKIN. Such compounds may include agonists, antagonists, or partial or inverse agonists. In one embodiment, an assay is performed under conditions permissive for PKIN activity, wherein PKIN is combined with at least one test compound, and the activity of PKIN in the presence of a test compound is compared with the activity of PKIN in the absence of the test compound. A change in the activity of PKIN in the presence of the test compound is indicative of a compound that modulates the activity of PKIN. Alternatively, a test compound is combined with an in vitro or cell-free system comprising PKIN under conditions suitable for PKIN activity, and the assay is performed. In either of these assays, a test compound which modulates the activity of PKIN may do so indirectly and need not come in direct contact with the test compound. At least one and up to a plurality of test compounds may be screened.

[0203] In another embodiment, polynucleotides encoding PKIN or their mammalian homologs may be "knocked out" in an animal model system using homologous recombination in embryonic stem (ES) cells. Such techniques are well known in the art and are useful for the generation of animal models of human disease. (See, e.g., U.S. Pat. No. 5,175,383 and U.S. Pat. No. 5,767,337.) For example, mouse ES cells, such as the mouse 129/SvJ cell line, are derived from the early mouse embryo and grown in culture. The ES cells are transformed with a vector containing the gene of interest disrupted by a marker gene, e.g., the neomycin phosphotransferase gene (neo; Capecchi, M. R. (1989) Science 244:1288-1292). The vector integrates into the corresponding region of the host genome by homologous recombination. Alternatively, homologous recombination takes place using the Cre-loxP system to knockout a gene of interest in a tissue- or developmental stage-specific manner (Marth, J. D. (1996) Clin. Invest. 97:1999-2002; Wagner, K. U. et al. (1997) Nucleic Acids Res. 25:4323-4330). Transformed ES cells are identified and microinjected into mouse cell blastocysts such as those from the C57BL/6 mouse strain. The blastocysts are surgically transferred to pseudopregnant dams, and the resulting chimeric progeny are genotyped and bred to produce heterozygous or homozygous strains. Transgenic animals thus generated may be tested with potential therapeutic or toxic agents.

[0204] Polynucleotides encoding PKIN may also be manipulated in vitro in ES cells derived from human blastocysts. Human ES cells have the potential to differentiate into at least eight separate cell lineages including endoderm, mesoderm, and ectodermal cell types. These cell lineages differentiate into, for example, neural cells, hematopoietic lineages, and cardiomyocytes (Thomson, J. A. et al. (1998) Science 282:1145-1147).

[0205] Polynucleotides encoding PKIN can also be used to create "knockin" humanized animals (pigs) or transgenic animals (mice or rats) to model human disease. With knockin technology, a region of a polynucleotide encoding PKIN is injected into animal ES cells, and the injected sequence integrates into the animal cell genome. Transformed cells are injected into blastulae, and the blastulae are implanted as described above. Transgenic progeny or inbred lines are studied and treated with potential pharmaceutical agents to obtain information on treatment of a human disease. Alternatively, a mammal inbred to overexpress PKIN, e.g., by secreting PKIN in its milk, may also serve as a convenient source of that protein (Janne, J. et al. (1998) Biotechnol. Annu. Rev. 4:55-74).

[0206] Therapeutics

[0207] Chemical and structural similarity, e.g., in the context of sequences and motifs, exists between regions of PKIN and human kinases. In addition, the expression of PKIN is closely associated with cancer, diseased, proliferative, cardiac, tumorous, and digestive tissues, degenerative diseases of the brain, suggesting that PKIN plays a role in necrotic disorders affecting the central nervous system, and neuronal tissues (e.g. brain and spinal cord, see Table 6). Therefore, PKIN appears to play a role in maintenance and potentially the neoplastic transformation of cells of the central nervous system, and in cancer, immune disorders, disorders affecting growth and development, cardiovascular diseases, and lipid disorders. In the treatment of disorders associated with increased PKIN expression or activity, it is desirable to decrease the expression or activity of PKIN. In the treatment of disorders associated with decreased PKIN expression or activity, it is desirable to increase the expression or activity of PKIN.

[0208] Therefore, in one embodiment, PKIN or a fragment or derivative thereof may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of PKIN. Examples of such disorders include, but are not limited to, a cancer, such as adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, and, in particular, cancers of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus, leukemias such as multiple myeloma and lymphomas such as Hodgkin's disease; an immune disorder, such as acquired immunodeficiency syndrome (AIDS), Addison's disease, adult respiratory distress syndrome, allergies, ankylosing spondylitis, amyloidosis, anemia, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmune thyroiditis, autoimmune polyendocrinopathy-candidiasis- -ectodermal dystrophy (APECED), bronchitis, cholecystitis, contact dermatitis, Crohn's disease, atopic dermatitis, dermatomyositis, diabetes mellitus, emphysema, episodic lymphopenia with lymphocytotoxins, erythroblastosis fetalis, erythema nodosum, atrophic gastritis, glomerulonephritis, Goodpasture's syndrome, gout, Graves' disease, Hashimoto's thyroiditis, hypereosinophilia, irritable bowel syndrome, multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, osteoarthritis, osteoporosis, pancreatitis, polymyositis, psoriasis, Reiter's syndrome, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic anaphylaxis, systemic lupus erytbematosus, systemic sclerosis, thrombocytopenic purpura, ulcerative colitis, uveitis, Werner syndrome, complications of cancer, hemodialysis, and extracorporeal circulation, viral, bacterial, fungal, parasitic, protozoal, and helminthic infections, and trauma; a growth and developmental disorder, such as actinic keratosis, arteriosclerosis, atherosclerosis, bursitis, cirrhosis, hepatitis, mixed connective tissue disease (MCTD), myelofibrosis, paroxysmal nocturnal hemoglobinuria, polycythemia vera, psoriasis, primary thrombocythemia, and cancers including adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, and, in particular, cancers of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus, renal tubular acidosis, anemia, Cushing's syndrome, achondroplastic dwarfism, Duchenne and Becker muscular dystrophy, epilepsy, gonadal dysgenesis, WAGR syndrome (Wilms' tumor, aniridia, genitourinary abnormalities, and mental retardation), Smith-Magenis syndrome, myelodysplastic syndrome, hereditary mucoepithelial dysplasia, hereditary keratodermas, hereditary neuropathies such as Charcot-Marie-Tooth disease and neurofibromatosis, hypothyroidism, hydrocephalus, seizure disorders such as Syndenham's chorea and cerebral palsy, spina bifida, anencephaly, craniorachischisis, congenital glaucoma, cataract, and sensorineural hearing loss; a cardiovascular disease, such as arterioyenous fistula, atherosclerosis, hypertension, vasculitis, Raynaud's disease, aneurysms, arterial dissections, varicose veins, thrombophlebitis and phlebothrombosis, vascular tumors, and complications of thrombolysis, balloon angioplasty, vascular replacement, and coronary artery bypass graft surgery, congestive heart failure, ischemic heart disease, angina pectoris, myocardial infarction, hypertensive heart disease, degenerative valvular heart disease, calcific aortic valve stenosis, congenitally bicuspid aortic valve, mitral annular calcification, mitral valve prolapse, rheumatic fever and rheumatic heart disease, infective endocarditis, nonbacterial thrombotic endocarditis, endocarditis of systemic lupus erythematosus, carcinoid heart disease, cardiomyopathy, myocarditis, pericarditis, neoplastic heart disease, congenital heart disease, and complications of cardiac transplantation, congenital lung anomalies, atelectasis, pulmonary congestion and edema, pulmonary embolism, pulmonary hemorrhage, pulmonary infarction, pulmonary hypertension, vascular sclerosis, obstructive pulmonary disease, restrictive pulmonary disease, chronic obstructive pulmonary disease, emphysema, chronic bronchitis, bronchial asthma, bronchiectasis, bacterial pneumonia, viral and mycoplasmal pneumonia, lung abscess, pulmonary tuberculosis, diffuse interstitial diseases, pneumoconioses, sarcoidosis, idiopathic pulmonary fibrosis, desquamative interstitial pneumonitis, hypersensitivity pneumonitis, pulmonary eosinophilia bronchiolitis obliterans-organizing pneumonia, diffuse pulmonary hemorrhage syndromes, Goodpasture's syndromes, idiopathic pulmonary hemosiderosis, pulmonary involvement in collagen-vascular disorders, pulmonary alveolar proteinosis, lung tumors, inflammatory and noninflammatory pleural effusions, pneumothorax, pleural tumors, drug-induced lung disease, radiation-induced lung disease, and complications of lung transplantation; and a lipid disorder such as fatty liver, cholestasis, primary biliary cirrhosis, carnitine deficiency, carnitine palmitoyltransferase deficiency, myoadenylate deaminase deficiency, hypertriglyceridemia, lipid storage disorders such Fabry's disease, Gaucher's disease, Niemann-Pick's disease, metachromatic leukodystrophy, adrenoleukodystrophy, GM.sub.2 gangliosidosis, and ceroid lipofuscinosis, abetalipoproteinemia, Tangier disease, hyperlipoproteinemia, diabetes mellitus, lipodystrophy, lipomatoses, acute panniculitis, disseminated fat necrosis, adiposis dolorosa, lipoid adrenal hyperplasia, minimal change disease, lipomas, atherosclerosis, hypercholesterolemia, hypercholesterolemia with hypertriglyceridemia, primary hypoalphalipoproteinemia, hypothyroidism, renal disease, liver disease, lecithin:cholesterol acyltransferase deficiency, cerebrotendinous xanthomatosis, sitosterolemia, hypocholesterolemia, Tay-Sachs disease, Sandhoff's disease, hyperlipidemia, hyperlipemia, lipid myopathies, and obesity.

[0209] In another embodiment, a vector capable of expressing PKIN or a fragment or derivative thereof may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of PKIN including, but not limited to, those described above.

[0210] In a further embodiment, a composition comprising a substantially purified PKIN in conjunction with a suitable pharmaceutical carrier may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of PKIN including, but not limited to, those provided above.

[0211] In still another embodiment, an agonist which modulates the activity of PKIN may be administered to a subject to treat or prevent a disorder associated with decreased expression or activity of PKEIN including, but not limited to, those listed above.

[0212] In a further embodiment, an antagonist of PKIN may be administered to a subject to treat or prevent a disorder associated with increased expression or activity of PKIN. Examples of such disorders include, but are not limited to, those cancers, immune disorders, disorders affecting growth and development, cardiovascular diseases, and lipid disorders described above. In one aspect, an antibody which specifically binds PKIN may be used directly as an antagonist or indirectly as a targeting or delivery mechanism for bringing a pharmaceutical agent to cells or tissues which express PKIN.

[0213] In an additional embodiment, a vector expressing the complement of the polynucleotide encoding PKIN may be administered to a subject to treat or prevent a disorder associated with increased expression or activity of PKIN including, but not limited to, those described above.

[0214] In other embodiments, any of the proteins, antagonists, antibodies, agonists, complementary sequences, or vectors of the invention may be administered in combination with other appropriate therapeutic agents. Selection of the appropriate agents for use in combination therapy may be made by one of ordinary skill in the art, according to conventional pharmaceutical principles. The combination of therapeutic agents may act synergistically to effect the treatment or prevention of the various disorders described above. Using this approach, one may be able to achieve therapeutic efficacy with lower dosages of each agent, thus reducing the potential for adverse side effects.

[0215] An antagonist of PKIN may be produced using methods which are generally known in the art. In particular, purified PKIN may be used to produce antibodies or to screen libraries of pharmaceutical agents to identify those which specifically bind PKIN. Antibodies to PKIN may also be generated using methods that are well known in the art. Such antibodies may include, but are not limited to, polyclonal, monoclonal, chimeric, and single chain antibodies, Fab fragments, and fragments produced by a Fab expression library. Neutralizing antibodies (i.e., those which inhibit dimer formation) are generally preferred for therapeutic use.

[0216] For the production of antibodies, various hosts including goats, rabbits, rats, mice, humans, and others may be immunized by injection with PKIN or with any fragment or oligopeptide thereof which has immunogenic properties. Depending on the host species, various adjuvants may be used to increase immunological response. Such adjuvants include, but are not limited to, Freund's, mineral gels such as aluminum hydroxide, and surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, KLH, and dinitrophenol. Among adjuvants used in humans, BCG (bacilli Calmette-Guerin) and Corynebacterium parvum are especially preferable.

[0217] It is preferred that the oligopeptides, peptides, or fragments used to induce antibodies to PKIN have an amino acid sequence consisting of at least about 5 amino acids, and generally will consist of at least about 10 amino acids. It is also preferable that these oligopeptides, peptides, or fragments are identical to a portion of the amino acid sequence of the natural protein. Short stretches of PKIN amino acids may be fused with those of another protein, such as KLH, and antibodies to the chimeric molecule may be produced.

[0218] Monoclonal antibodies to PKIN may be prepared using any technique which provides for the production of antibody molecules by continuous cell lines in culture. These include, but are not limited to, the hybridoma technique, the human B-cell hybridoma technique, and the EBV-hybridoma technique. (See, e.g., Kohler, G. et al. (1975) Nature 256:495497; Kozbor, D. et al. (1985) J. Immunol. Methods 81:31-42; Cote, R. J. et al. (1983) Proc. Natl. Acad. Sci. USA 80:2026-2030; and Cole, S. P. et al. (1984) Mol. Cell Biol. 62:109-120.)

[0219] In addition, techniques developed for the production of "chimeric antibodies," such as the splicing of mouse antibody genes to human antibody genes to obtain a molecule with appropriate antigen specificity and biological activity, can be used. (See, e.g., Morrison, S. L. et al. (1984) Proc. Natl. Acad. Sci. USA 81:6851-6855; Neuberger, M. S. et al. (1984) Nature 312:604-608; and Takeda, S. et al. (1985) Nature 314:452-454.) Alternatively, techniques described for the production of single chain antibodies may be adapted, using methods known in the art, to produce PKIN-specific single chain antibodies. Antibodies with related specificity, but of distinct idiotypic composition, may be generated by chain shuffling from random combinatorial immunoglobulin libraries. (See, e.g., Burton, D. R. (1991) Proc. Natl. Acad. Sci. USA 88:10134-10137.) Antibodies may also be produced by inducing in vivo production in the lymphocyte population or by screening immunoglobulin libraries or panels of highly specific binding reagents as disclosed in the literature. (See, e.g., Orlandi, R. et al. (1989) Proc. Natl. Acad. Sci. USA 86:3833-3837; Winter, G. et al. (1991) Nature 349:293-299.) Antibody fragments which contain specific binding sites for PKIN may also be generated. For example, such fragments include, but are not limited to, F(ab').sub.2 fragments produced by pepsin digestion of the antibody molecule and Fab fragments generated by reducing the disulfide bridges of the F(ab')2 fragments. Alternatively, Fab expression libraries may be constructed to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity. (See, e.g., Huse, W. D. et al. (1989) Science 246:1275-1281.) Various immunoassays may be used for screening to identify antibodies having the desired specificity. Numerous protocols for competitive binding or immunoradiometric assays using either polyclonal or monoclonal antibodies with established specificities are well known in the art. Such immunoassays typically involve the measurement of complex formation between PKIN and its specific antibody. A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering PKIN epitopes is generally used, but a competitive binding assay may also be employed (Pound, supra).

[0220] Various methods such as Scatchard analysis in conjunction with radioimmunoassay techniques may be used to assess the affinity of antibodies for PKIN. Affinity is expressed as an association constant, K.sub.a, which is defined as the molar concentration of PKIN-antibody complex divided by the molar concentrations of free antigen and free antibody under equilibrium conditions. The K.sub.a determined for a preparation of polyclonal antibodies, which are heterogeneous in their affinities for multiple PKIN epitopes, represents the average affinity, or avidity, of the antibodies for PKIN. The K.sub.a determined for a preparation of monoclonal antibodies, which are monospecific for a particular PKIN epitope, represents a true measure of affinity. High-affinity antibody preparations with K.sub.a ranging from about 10.sup.9 to 10.sup.12 L/mole are preferred for use in immunoassays in which the PKIN-antibody complex must withstand rigorous manipulations. Low-affinity antibody preparations with K, ranging from about 10.sup.6 to 10.sup.7 L/mole are preferred for use in immunopurification and similar procedures which ultimately require dissociation of PKIN, preferably in active form, from the antibody (Catty, D. (1988) Antibodies. Volume I: A Practical Approach, IRL Press, Washington D.C.; Liddell, J. E. and A. Cryer (1991) A Practical Guide to Monoclonal Antibodies, John Wiley & Sons, New York N.Y.).

[0221] The titer and avidity of polyclonal antibody preparations may be further evaluated to determine the quality and suitability of such preparations for certain downstream applications. For example, a polyclonal antibody preparation containing at least 1-2 mg specific antibody/ml, preferably 5-10 mg specific antibody/ml, is generally employed in procedures requiring precipitation of PKIN-antibody complexes. Procedures for evaluating antibody specificity, titer, and avidity, and guidelines for antibody quality and usage in various applications, are generally available. (See, e.g., Catty, supra, and Coligan et al. supra.) In another embodiment of the invention, the polynucleotides encoding PKIN, or any fragment or complement thereof, may be used for therapeutic purposes. In one aspect, modifications of gene expression can be achieved by designing complementary sequences or antisense molecules (DNA, RNA, PNA, or modified oligonucleotides) to the coding or regulatory regions of the gene encoding PKIN. Such technology is well known in the art, and antisense oligonucleotides or larger fragments can be designed from various locations along the coding or control regions of sequences encoding PKIN. (See, e.g., Agrawal, S., ed. (1996) Antisense Therapeutics, Humana Press Inc., Totawa N.J.)

[0222] In therapeutic use, any gene delivery system suitable for introduction of the antisense sequences into appropriate target cells can be used. Antisense sequences can be delivered intracellularly in the form of an expression plasmid which, upon transcription, produces a sequence complementary to at least a portion of the cellular sequence encoding the target protein. (See, e.g., Slater, J. E. et al. (1998) J. Allergy Cli. Immunol. 102(3):469-475; and Scanlon, K. J. et al. (1995) 9(13):1288-1296.) Antisense sequences can also be introduced intracellularly through the use of viral vectors, such as retrovirus and adeno-associated virus vectors. (See, e.g., Miller, A. D. (1990) Blood 76:271; Ausubel, supra; Uckert, W. and W. Walther (1994) Pharmacol. Ther. 63(3):323-347.) Other gene delivery mechanisms include liposome-derived systems, artificial viral envelopes, and other systems known in the art. (See, e.g., Rossi, J. J. (1995) Br. Med. Bull. 51(1):217-225; Boado, R. J. et al. (1998) J. Pharm. Sci. 87(11):1308-1315; and Morris, M. C. et al. (1997) Nucleic Acids Res. 25(14):2730-2736.)

[0223] In another embodiment of the invention, polynucleotides encoding PKIN may be used for somatic or germline gene therapy. Gene therapy may be performed to (i) correct a genetic deficiency (e.g., in the cases of severe combined immunodeficiency (SCID)-X1 disease characterized by X-linked inheritance (Cavazzana-Calvo, M. et al. (2000) Science 288:669-672), severe combined immunodeficiency syndrome associated with an inherited adenosine deaminase (ADA) deficiency (Blaese, R. M. et al. (1995) Science 270:475-480; Bordignon, C. et al. (1995) Science 270:470-475), cystic fibrosis (Zabner, J. et al. (1993) Cell 75:207-216; Crystal, R. G. et al. (1995) Hum Gene Therapy 6:643-666; Crystal, R. G. et al. (1995) Hum. Gene Therapy 6:667-703), thalassamias, familial hypercholesterolemia, and hemophilia resulting from Factor VIII or Factor IX deficiencies (Crystal, R. G. (1995) Science 270:404410; Verma, I. M. and N. Somia (1997) Nature 389:239-242)), (ii) express a conditionally lethal gene product (e.g., in the case of cancers which result from unregulated cell proliferation), or (iii) express a protein which affords protection against intracellular parasites (e.g., against human retroviruses, such as human immunodeficiency virus (HIV) (Baltimore, D. (1988) Nature 335:395-396; Poeschla, E. et al. (1996) Proc. Natl. Acad. Sci. USA. 93:11395-11399), hepatitis B or C virus (HBV, HCV); fungal parasites, such as Candida albicans and Paracoccidioides brasiliensis; and protozoan parasites such as Plasmodium falciparum and Trypanosoma cruzi). In the case where a genetic deficiency in PKIN expression or regulation causes disease, the expression of PKIN from an appropriate population of transduced cells may alleviate the clinical manifestations caused by the genetic deficiency.

[0224] In a further embodiment of the invention, diseases or disorders caused by deficiencies in PKIN are treated by constructing mammalian expression vectors encoding PKIN and introducing these vectors by mechanical means into PKIN-deficient cells. Mechanical transfer technologies for use with cells in vivo or ex vitro include (i) direct DNA microinjection into individual cells, (ii) ballistic gold particle delivery, (iii) liposome-mediated transfection, (iv) receptor-mediated gene transfer, and (v) the use of DNA transposons (Morgan, R. A. and W. F. Anderson (1993) Annu. Rev. Biochem. 62:191-217; Ivics, Z. (1997) Cell 91:501-510; Boulay, J -L. and H. Rcipon (1998) Curr. Opin. Biotechnol. 9:445-450).

[0225] Expression vectors that may be effective for the expression of PKIN include, but are not limited to, the PcDNA 3.1, EPITAG, PRCCMV2, PREP, PVAX vectors (Invitrogen, Carlsbad Calif.), PCMV-SCRIPT, PCMV-TAG, PEGSH/PERV (Stratagene, La Jolla Calif.), and PTET-OFF, PTET-ON, PTRE2, PTRE2-LUC, PTK-HYG (Clontech, Palo Alto Calif.). PKN may be expressed using (i) a constitutively active promoter, (e.g., from cytomegalovirus (CMV), Rous sarcoma virus (RSV), SV40 virus, thymidine kinase (TK), or mactin genes), (ii) an inducible promoter (e.g., the tetracycline-regulated promoter (Gossen, M. and H. Bujard (1992) Proc. Natl. Acad. Sci. USA 89:5547-5551, Gossen, M. et al. (1995) Science 268:1766-1769; Rossi, F. M. V. and H. M. Blau (1998) Curr. Opin. Biotechnol. 9:451-456), commercially available in the T-REX plasmid (Invitrogen)); the ecdysone-inducible promoter (available in the plasmids PVGRXR and PIND; Invitrogen); the FKS06/rapamycin inducible promoter; or the RU486/mifepristone inducible promoter (Rossi, F. M. V. and Blau, H. M. supra)), or (iii) a tissue-specific promoter or the native promoter of the endogenous gene encoding PKIN from a normal individual.

[0226] Commercially available liposome transformation kits (e.g., the PERFECT LIPID TRANSFECTION KIT, available from Invitrogen) allow one with ordinary skill in the art to deliver polynucleotides to target cells in culture and require minimal effort to optimize experimental parameters. In the alternative, transformation is performed using the calcium phosphate method (Graham, F. L. and A. J. Eb (1973) Virology 52:456467), or by electroporation (Neumann, E. et al. (1982) EMBO J. 1:841-845). The introduction of DNA to primary cells requires modification of these standardized mammalian transfection protocols.

[0227] In another embodiment of the invention, diseases or disorders caused by genetic defects with respect to PKN expression are treated by constructing a retrovirus vector consisting of (i) the polynucleotide encoding PKIN under the control of an independent promoter or the retrovirus long terminal repeat (LTR) promoter, (ii) appropriate RNA packaging signals, and (iii) a Rev-responsive element (RRE) along with additional retrovirus cis-acting RNA sequences and coding sequences required for efficient vector propagation. Retrovirus vectors (e.g., PFB and PFBNEO) are commercially available (Stratagene) and are based on published data (Riviere, I. et al. (1995) Proc. Natl. Acad. Sci. USA 92:6733-6737), incorporated by reference herein. The vector is propagated in an appropriate vector producing cell line (VPCL) that expresses an envelope gene with a tropism for receptors on the target cells or a promiscuous envelope protein such as VSVg (Armentano, D. et al. (1987) J. Virol. 61:1647-1650; Bender, M. A. et al. (1987) J. Virol. 61:1639-1646; Adam, M. A. and A. D. Miller (1988) J. Virol. 62:3802-3806; Dull, T. et al. (1998) J. Virol. 72:8463-8471; Zufferey, R. et al. (1998) J. Virol. 72:9873-9880). U.S. Pat. No. 5,910,434 to Rigg ("Method for obtaining retrovirus packaging cell lines producing high transducing efficiency retroviral supernatant") discloses a method for obtaining retrovirus packaging cell lines and is hereby incorporated by reference. Propagation of retrovirus vectors, transduction of a population of cells (e.g., CD4.sup.+ T-cells), and the return of transduced cells to a patient are procedures well known to persons skilled in the art of gene therapy and have been well documented (Ranga, U. et al. (1997) J. Virol. 71:7020-7029; Bauer, G. et al. (1997) Blood 89:2259-2267; Bonyhadi, M. L. (1997) J. Virol. 71:4707-4716; Ranga, U. et al. (1998) Proc. Natl. Acad. Sci. USA 95:1201-1206; Su, L. (1997) Blood 89:2283-2290).

[0228] In the alternative, an adenovirus-based gene therapy delivery system is used to deliver polynucleotides encoding PKIN to cells which have one or more genetic abnormalities with respect to the expression of PKIN. The construction and packaging of adenovirus-based vectors are well known to those with ordinary skill in the art. Replication defective adenovirus vectors have proven to be versatile for importing genes encoding immunoregulatory proteins into intact islets in the pancreas (Csete, M. E. et al. (1995) Transplantation 27:263-268). Potentially useful adenoviral vectors are described in U.S. Pat. No. 5,707,618 to Armentano ("Adenovirus vectors for gene therapy"), hereby incorporated by reference. For adenoviral vectors, see also Antinozzi, P. A. et al. (1999) Annu. Rev. Nutr. 19:511-544 and Verma, I. M. and N. Somia (1997) Nature 18:389:239-242, both incorporated by reference herein.

[0229] In another alternative, a herpes-based, gene therapy delivery system is used to deliver polynucleotides encoding PKIN to target cells which have one or more genetic abnormalities with respect to the expression of PKIN. The use of herpes simplex virus (HSV)-based vectors may be especially valuable for introducing PKIN to cells of the central nervous system, for which HSV has a tropism. The construction and packaging of herpes-based vectors are well known to those with ordinary skill in the art. A replication-competent herpes simplex virus (HSV) type 1-based vector has been used to deliver a reporter gene to the eyes of primates (Liu, X. et al. (1999) Exp. Eye Res. 169:385-395). The construction of a HSV-1 virus vector has also been disclosed in detail in U.S. Pat. No. 5,804,413 to DeLuca ("Herpes simplex virus strains for gene transfer"), which is hereby incorporated by reference. U.S. Pat. No. 5,804,413 teaches the use of recombinant HSV d92 which consists of a genome containing at least one exogenous gene to be transferred to a cell under the control of the appropriate promoter for purposes including human gene therapy. Also taught by this patent are the construction and use of recombinant HSV strains deleted for ICP4, ICP27 and ICP22. For HSV vectors, see also Goins, W. F. et al. (1999) J. Virol. 73:519-532 and Xu, H. et al. (1994) Dev. Biol. 163:152-161, hereby incorporated by reference. The manipulation of cloned herpesvirus sequences, the generation of recombinant virus following the transfection of multiple plasmids containing different segments of the large herpesvirus genomes, the growth and propagation of herpesvirus, and the infection of cells with herpesvirus are techniques well known to those of ordinary skill in the art.

[0230] In another alternative, an alphavirus (positive, single-stranded RNA virus) vector is used to deliver polynucleotides encoding PKIN to target cells. The biology of the prototypic alphavirus, Semliki Forest Virus (SFV), has been studied extensively and gene transfer vectors have been based on the SFV genome (Garoff, H. and K. -J. Li (1998) Curr. Opin. Biotechnol. 9:464-469). During alphavirus RNA replication, a subgenomic RNA is generated that normally encodes the viral capsid proteins. This subgenomic RNA replicates to higher levels than the full length genomic RNA, resulting in the overproduction of capsid proteins relative to the viral proteins with enzymatic activity (e.g., protease and polymerase). Similarly, inserting the coding sequence for PKIN into the alphavirus genome in place of the capsid-coding region results in the production of a large number of PKIN-coding RNAs and the synthesis of high levels of PKIN in vector transduced cells. While alphavirus infection is typically associated with cell lysis within a few days, the ability to establish a persistent infection in hamster normal kidney cells (BHK-21) with a variant of Sindbis virus (SIN) indicates that the lytic replication of alphaviruses can be altered to suit the needs of the gene therapy application (Dryga, S. A. et al. (1997) Virology 228:74-83). The wide host range of alphaviruses will allow the introduction of PKIN into a variety of cell types. The specific transduction of a subset of cells in a population may require the sorting of cells prior to transduction. The methods of manipulating infectious cDNA clones of alphaviruses, performing alphavirus cDNA and RNA transfections, and performing alphavirus infections, are well known to those with ordinary skill in the art.

[0231] Oligonucleotides derived from the transcription initiation site, e.g., between about positions -10 and +10 from the start site, may also be employed to inhibit gene expression. Similarly, inhibition can be achieved using triple helix base-pairing methodology. Triple helix pairing is useful because it causes inhibition of the ability of the double helix to open sufficiently for the binding of polymerases, transcription factors, or regulatory molecules. Recent therapeutic advances using triplex DNA have been described in the literature. (See, e.g., Gee, J. E. et al. (1994) in Huber, B. E. and B. I. Carr, Molecular and Immunologic Approaches, Futura Publishing, Mt. Kisco N.Y., pp. 163-177.) A complementary sequence or antisense molecule may also be designed to block translation of mRNA by preventing the transcript from binding to ribosomes.

[0232] Ribozymes, enzymatic RNA molecules, may also be used to catalyze the specific cleavage of RNA. The mechanism of ribozyme action involves sequence-specific hybridization of the ribozyme molecule to complementary target RNA, followed by endonucleolytic cleavage. For example, engineered hammerhead motif ribozyme molecules may specifically and efficiently catalyze endonucleolytic cleavage of sequences encoding PKIN.

[0233] Specific ribozyme cleavage sites within any potential RNA target are initially identified by scanning the target molecule for ribozyme cleavage sites, including the following sequences: GUA, GUU, and GUC. Once identified, short RNA sequences of between 15 and 20 ribonucleotides, corresponding to the region of the target gene containing the cleavage site, may be evaluated for secondary structural features which may render the oligonucleotide inoperable. The suitability of candidate targets may also be evaluated by testing accessibility to hybridization with complementary oligonucleotides using ribonuclease protection assays.

[0234] Complementary ribonucleic acid molecules and ribozymes of the invention may be prepared by any method known in the art for the synthesis of nucleic acid molecules. These include techniques for chemically synthesizing oligonucleotides such as solid phase phosphoramidite chemical synthesis. Alternatively, RNA molecules may be generated by in vitro and in vivo transcription of DNA sequences encoding PKIN. Such DNA sequences may be incorporated into a wide variety of vectors with suitable RNA polymerase promoters such as T7 or SP6. Alternatively, these cDNA constructs that synthesize complementary RNA, constitutively or inducibly, can be introduced into cell lines, cells, or tissues.

[0235] RNA molecules may be modified to increase intracellular stability and half-life. Possible modifications include, but are not limited to, the addition of flanking sequences at the 5' and/or 3' ends of the molecule, or the use of phosphorothioate or 2'O-methyl rather than phosphodiesterase linkages within the backbone of the molecule. This concept is inherent in the production of PNAs and can be extended in all of these molecules by the inclusion of nontraditional bases such as inosine, queosine, and wybutosine, as well as acetyl-, methyl-, thio-, and similarly modified forms of adenine, cytidine, guanine, thymine, and uridine which are not as easily, recognized by endogenous endonucleases.

[0236] An additional embodiment of the invention encompasses a method for screening for a compound which is effective in altering expression of a polynucleotide encoding PKIN. Compounds which may be effective in altering expression of a specific polynucleotide may include, but are not limited to, oligonucleotides, antisense oligonucleotides, triple helix-forming oligonucleotides, transcription factors and other polypeptide transcriptional regulators, and non-macromolecular chemical entities which are capable of interacting with specific polynucleotide sequences. Effective compounds may alter polynucleotide expression by acting as either inhibitors or promoters of polynucleotide expression. Thus, in the treatment of disorders associated with increased PKIN expression or activity, a compound which specifically inhibits expression of the polynucleotide encoding PKIN may be therapeutically useful, and in the treament of disorders associated with decreased PKIN expression or activity, a compound which specifically promotes expression of the polynucleotide encoding PKIN may be therapeutically useful.

[0237] At least one, and up to a plurality, of test compounds may be screened for effectiveness in altering expression of a specific polynucleotide. A test compound may be obtained by any method commonly known in the art, including chemical modification of a compound known to be effective in altering polynucleotide expression; selection from an existing, commercially-available or proprietary library of naturally-occurring or non-natural chemical compounds; rational design of a compound based on chemical and/or structural properties of the target polynucleotide; and selection from a library of chemical compounds created combinatorially or randomly. A sample comprising a polynucleotide encoding PKIN is exposed to at least one test compound thus obtained. The sample may comprise, for example, an intact or permeabilized cell, or an in vitro cell-free or reconstituted biochemical system. Alterations in the expression of a polynucleotide encoding PKIN are assayed by any method commonly known in the art. Typically, the expression of a specific nucleotide is detected by hybridization with a probe having a nucleotide sequence complementary to the sequence of the polynucleotide encoding PKIN. The amount of hybridization may be quantified, thus forming the basis for a comparison of the expression of the polynucleotide both with and without exposure to one or more test compounds. Detection of a change in the expression of a polynucleotide exposed to a test compound indicates that the test compound is effective in altering the expression of the polynucleotide. A screen for a compound effective in altering expression of a specific polynucleotide can be carried out, for example, using a Schizosaccharomyces pombe gene expression system (Atkins, D. et al. (1999) U.S. Pat. No. 5,932,435; Arndt, G. M. et al. (2000) Nucleic Acids Res. 28:E15) or a human cell line such as HeLa cell (Clarke, M. L. et al. (2000) Biochem. Biophys. Res. Commun. 268:8-13). A particular embodiment of the present invention involves screening a combinatorial library of oligonucleotides (such as deoxyribonucleotides, ribonucleotides, peptide nucleic acids, and modified oligonucleotides) for antisense activity against a specific polynucleotide sequence (Bruice, T. W. et al. (1997) U.S. Pat. No. 5,686,242; Bruice, T. W. et al. (2000) U.S. Pat. No. 6,022,691).

[0238] Many methods for introducing vectors into cells or tissues are available and equally suitable for use in vivo, in vitro, and ex vivo. For ex vivo therapy, vectors may be introduced into stem cells taken from the patient and clonally propagated for autologous transplant back into that same patient. Delivery by transfection, by liposome injections, or by polycationic amino polymers may be achieved using methods which are well known in the art. (See, e.g., Goldman, C. K. et al. (1997) Nat. Biotechnol. 15:462466.)

[0239] Any of the therapeutic methods described above may be applied to any subject in need of such therapy, including, for example, mammals such as humans, dogs, cats, cows, horses, rabbits, and monkeys.

[0240] An additional embodiment of the invention relates to the administration of a composition which generally comprises an active ingredient formulated with a pharmaceutically acceptable excipient. Excipients may include, for example, sugars, starches, celluloses, gums, and proteins. Various formulations are commonly known and are thoroughly discussed in the latest edition of Remington's Pharmaceutical Sciences (Maack Publishing, Easton Pa.). Such compositions may consist of PKIN, antibodies to PKIN, and mimetics, agonists, antagonists, or inhibitors of PKIN.

[0241] The compositions utilized in this invention may be administered by any number of routes including, but not limited to, oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, intraventricular, pulmonary, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal means.

[0242] Compositions for pulmonary administration may be prepared in liquid or dry powder form. These compositions are generally aerosolized immediately prior to inhalation by the patient. In the case of small molecules (e.g. traditional low molecular weight organic drugs), aerosol delivery of fast-acting formulations is well-known in the art. In the case of macromolecules (e.g. larger peptides and proteins), recent developments in the field of pulmonary delivery via the alveolar region of the lung have enabled the practical delivery of drugs such as insulin to blood circulation (see, e.g., Patton, J. S. et al., U.S. Pat. No. 5,997,848). Pulmonary delivery has the advantage of administration without needle injection, and obviates the need for potentially toxic penetration enhancers.

[0243] Compositions suitable for use in the invention include compositions wherein the active ingredients are contained in an effective amount to achieve the intended purpose. The determination of an effective dose is well within the capability of those skilled in the art.

[0244] Specialized forms of compositions may be prepared for direct intracellular delivery of macromolecules comprising PKIN or fragments thereof. For example, liposome preparations containing a cell-impermeable macromolecule may promote cell fusion and intracellular delivery of the macromolecule. Alternatively, PKIN or a fragment thereof may be joined to a short cationic N-terminal portion from the HIV Tat-1 protein. Fusion proteins thus generated have been found to transduce into the cells of all tissues, including the brain, in a mouse model system (Schwarze, S. R. et al. (1999) Science 285:1569-1572).

[0245] For any compound, the therapeutically effective dose can be estimated initially either in cell culture assays, e.g., of neoplastic cells, or in animal models such as mice, rats, rabbits, dogs, monkeys, or pigs. An animal model may also be used to determine the appropriate concentration range and route of administration. Such information can then be used to determine useful doses and routes for administration in humans.

[0246] A therapeutically effective dose refers to that amount of active ingredient, for example PKIN or fragments thereof, antibodies of PKIN, and agonists, antagonists or inhibitors of PKIN, which ameliorates the symptoms or condition. Therapeutic efficacy and toxicity may be determined by standard pharmaceutical procedures in cell cultures or with experimental animals, such as by calculating the ED.sub.50 (the dose therapeutically effective in 50% of the population) or LD.sub.50 (the dose lethal to 50% of the population) statistics. The dose ratio of toxic to therapeutic effects is the therapeutic index, which can be expressed as the LD.sub.50/ED.sub.50 ratio. Compositions which exhibit large therapeutic indices are preferred. The data obtained from cell culture assays and animal studies are used to formulate a range of dosage for human use. The dosage contained in such compositions is preferably within a range of circulating concentrations that includes the ED.sub.50 with little or no toxicity. The dosage varies within this range depending upon the dosage form employed, the sensitivity of the patient, and the route of administration.

[0247] The exact dosage will be determined by the practitioner, in light of factors related to the subject requiring treatment. Dosage and administration are adjusted to provide sufficient levels of the active moiety or to maintain the desired effect. Factors which may be taken into account include the severity of the disease state, the general health of the subject, the age, weight, and gender of the subject, time and frequency of administration, drug combination(s), reaction sensitivities, and response to therapy. Long-acting compositions may be administered every 3 to 4 days, every week, or biweekly depending on the half-life and clearance rate of the particular formulation.

[0248] Normal dosage amounts may vary from about 0.1 .mu.g to 100,000 .mu.g, up to a total dose of about 1 gram, depending upon the route of administration. Guidance as to particular dosages and methods of delivery is provided in the literature and generally available to practitioners in the art. Those skilled in the art will employ different formulations for nucleotides than for proteins or their inhibitors. Similarly, delivery of polynucleotides or polypeptides will be specific to particular cells, conditions, locations, etc.

[0249] Diagnostics

[0250] In another embodiment, antibodies which specifically bind PKIN may be used for the diagnosis of disorders characterized by expression of PKIN, or in assays to monitor patients being treated with PKIN or agonists, antagonists, or inhibitors of PKIN. Antibodies useful for diagnostic purposes may be prepared in the same manner as described above for therapeutics. Diagnostic assays for PKIN include methods which utilize the antibody and a label to detect PKIN in human body fluids or in extracts of cells or tissues. The antibodies may be used with or without modification, and may be labeled by covalent or non-covalent attachment of a reporter molecule. A wide variety of reporter molecules, several of which are described above, are known in the art and may be used.

[0251] A variety of protocols for measuring PKIN, including ELISAs, RIAs, and FACS, are known in the art and provide a basis for diagnosing altered or abnormal levels of PKIN expression. Normal or standard values for PKIN expression are established by combining body fluids or cell extracts taken from normal mammalian subjects, for example, human subjects, with antibodies to PKIN under conditions suitable for complex formation. The amount of standard complex formation may be quantitated by various methods, such as photometric means. Quantities of PKIN expressed in subject, control, and disease samples from biopsied tissues are compared with the standard values. Deviation between standard and subject values establishes the parameters for diagnosing disease.

[0252] In another embodiment of the invention, the polynucleotides encoding PKIN may be used for diagnostic purposes. The polynucleotides which may be used include oligonucleotide sequences, complementary RNA and DNA molecules, and PNAs. The polynucleotides may be used to detect and quantify gene expression in biopsied tissues in which expression of PKIN may be correlated with disease. The diagnostic assay may be used to determine absence, presence, and excess expression of PKIN, and to monitor regulation of PKIN levels during therapeutic intervention.

[0253] In one aspect, hybridization with PCR probes which are capable of detecting polynucleotide sequences, including genomic sequences, encoding PKIN or closely related molecules may be used to identify nucleic acid sequences which encode PKIN. The specificity of the probe, whether it is made from a highly specific region, e.g., the 5' regulatory region, or from a less specific region, e.g., a conserved motif, and the stringency of the hybridization or amplification will determine whether the probe identifies only naturally occurring sequences encoding PKIN, allelic variants, or related sequences.

[0254] Probes may also be used for the detection of related sequences, and may have at least 50% sequence identity to any of the PKIN encoding sequences. The hybridization probes of the subject invention may be DNA or RNA and may be derived from the sequence of SEQ ID NO:19-36 or from genomic sequences including promoters, enhancers, and introns of the PKIN gene.

[0255] Means for producing specific hybridization probes for DNAs encoding PKIN include the cloning of polynucleotide sequences encoding PKIN or PKIN derivatives into vectors for the production of mRNA probes. Such vectors are known in the art, are commercially available, and may be used to synthesize RNA probes in vitro by means of the addition of the appropriate RNA polymerases and the appropriate labeled nucleotides. Hybridization probes may be labeled by a variety of reporter groups, for example, by radionuclides such as .sup.32P or .sup.35S, or by enzymatic labels, such as alkaline phosphatase coupled to the probe via avidin/biotin coupling systems, and the like.

[0256] Polynucleotide sequences encoding PKIN may be used for the diagnosis of disorders associated with expression of PKIN. Examples of such disorders include, but are not limited to, a cancer, such as adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, and, in particular, cancers of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus, leukemias such as multiple myeloma and lymphomas such as Hodgkin's disease; an immune disorder, such as acquired immunodeficiency syndrome (AIDS), Addison's disease, adult respiratory distress syndrome, allergies, ankylosing spondylitis, amyloidosis, anemia, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmune thyroiditis, autoimmune polyendocrinopathy-candidiasis- -ectodermal dystrophy (APECED), bronchitis, cholecystitis, contact dermatitis, Crohn's disease, atopic dermatitis, dermatomyositis, diabetes mellitus, emphysema, episodic lymphopenia with lymphocytotoxins, erythroblastosis fetalis, erythema nodosum, atrophic gastritis, glomerulonephritis, Goodpasture's syndrome, gout, Graves' disease, Hashimoto's thyroiditis, hypereosinophilia, irritable bowel syndrome, multiple sclerosis, myasthenia gravis, myocardial or pericardial inflammation, osteoarthritis, osteoporosis, pancreatitis, polymyositis, psoriasis, Reiter's syndrome, rheumatoid arthritis, scleroderma, Sjogren's syndrome, systemic anaphylaxis, systemic lupus erythematosus, systemic sclerosis, thrombocytopenic purpura, ulcerative colitis, uveitis, Werner syndrome, complications of cancer, hemodialysis, and extracorporeal circulation, viral, bacterial, fungal, parasitic, protozoal, and helminthic infections, and trauma; a growth and developmental disorder, such as actinic keratosis, arteriosclerosis, atherosclerosis, bursitis, cirrhosis, hepatitis, mixed connective tissue disease (MCID), myelofibrosis, paroxysmal nocturnal hemoglobinuria, polycythemia vera, psoriasis, primary thrombocythemia, and cancers including adenocarcinoma, leukemia, lymphoma, melanoma, myeloma, sarcoma, teratocarcinoma, and, in particular, cancers of the adrenal gland, bladder, bone, bone marrow, brain, breast, cervix, gall bladder, ganglia, gastrointestinal tract, heart, kidney, liver, lung, muscle, ovary, pancreas, parathyroid, penis, prostate, salivary glands, skin, spleen, testis, thymus, thyroid, and uterus, renal tubular acidosis, anemia, Cushing's syndrome, achondroplastic dwarfism, Duchenne and Becker muscular dystrophy, epilepsy, gonadal dysgenesis, WAGR syndrome (Wilms' tumor, aniridia, genitourinary abnormalities, and mental retardation), Smith-Magenis syndrome, myelodysplastic syndrome, hereditary mucoepithelial dysplasia, hereditary keratodermas, hereditary neuropathies such as Charcot-Marie-Tooth disease and neurofibromatosis, hypothyroidism, hydrocephalus, seizure disorders such as Syndenham's chorea and cerebral palsy, spina bifida, anencephaly, craniorachischisis, congenital glaucoma, cataract, and sensorineural hearing loss; a cardiovascular disease, such as arterioyenous fistula, atherosclerosis, hypertension, vasculitis, Raynaud's disease, aneurysms, arterial dissections, varicose veins, thrombophlebitis and phlebothrombosis, vascular tumors, and complications of thrombolysis, balloon angioplasty, vascular replacement, and coronary artery bypass graft surgery, congestive heart failure, ischemic heart disease, angina pectoris, myocardial infarction, hypertensive heart disease, degenerative valvular heart disease, calcific aortic valve stenosis, congenitally bicuspid aortic valve, mitral annular calcification, mitral valve prolapse, rheumatic fever and rheumatic heart disease, infective endocarditis, nonbacterial thrombotic endocarditis, endocarditis of systemic lupus erythematosus, carcinoid heart disease, cardiomyopathy, myocarditis, pericarditis, neoplastic heart disease, congenital heart disease, and complications of cardiac transplantation, congenital lung anomalies, atelectasis, pulmonary congestion and edema, pulmonary embolism, pulmonary hemorrhage, pulmonary infarction, pulmonary hypertension, vascular sclerosis, obstructive pulmonary disease, restrictive pulmonary disease, chronic obstructive pulmonary disease, emphysema, chronic bronchitis, bronchial asthma, bronchiectasis, bacterial pneumonia, viral and mycoplasmal pneumonia, lung abscess, pulmonary tuberculosis, diffuse interstitial diseases, pneumocomoses, sarcoidosis, idiopathic pulmonary fibrosis, desquamative interstitial pneumonitis, hypersensitivity pneumonitis, pulmonary eosinophilia bronchiolitis obliterans-organizing pneumonia, diffuse pulmonary hemorrhage syndromes, Goodpasture's syndromes, idiopathic pulmonary hemosiderosis, pulmonary involvement in collagen-vascular disorders, pulmonary alveolar proteinosis, lung tumors, inflammatory and noninflammatory pleural effusions, pneumothorax, pleural tumors, drug-induced lung disease, radiation-induced lung disease, and complications of lung transplantation; and a lipid disorder such as fatty liver, cholestasis, primary biliary cirrhosis, carnitine deficiency, carnitine palmitoyltransferase deficiency, myoadenylate deaminase deficiency, hypertriglyceridemia, lipid storage disorders such Fabry's disease, Gaucher's disease, Niemann-Pick's disease, metachromatic leukodystrophy, adrenoleukodystrophy, GM.sub.2 gangliosidosis, and ceroid lipofuscinosis, abetalipoproteinemia, Tangier disease, hyperlipoproteinemia, diabetes mellitus, lipodystrophy, lipomatoses, acute panniculitis, disseminated fat necrosis, adiposis dolorosa, lipoid adrenal hyperplasia, minimal change disease, lipomas, atherosclerosis, hypercholesterolemia, hypercholesterolemia with hypertriglyceridemia, primary hypoalphalipoproteinemia, hypothyroidism, renal disease, liver disease, lecithin:cholesterol acyltransferase deficiency, cerebrotendinous xanthomatosis, sitosterolemia, hypocholesterolemia, Tay-Sachs disease, Sandhoff's disease, hyperlipidemia, hyperlipemia, lipid myopathies, and obesity. The polynucleotide sequences encoding PKIN may be used in Southern or northern analysis, dot blot, or other membrane-based technologies; in PCR technologies; in dipstick, pin, and multiformat ELISA-like assays; and in microarrays utilizing fluids or tissues from patients to detect altered PKIN expression. Such qualitative or quantitative methods are well known in the art.

[0257] In a particular aspect, the nucleotide sequences encoding PKIN may be useful in assays that detect the presence of associated disorders, particularly those mentioned above. The nucleotide sequences encoding PKIN may be labeled by standard methods and added to a fluid or tissue sample from a patient under conditions suitable for the formation of hybridization complexes. After a suitable incubation period, the sample is washed and the signal is quantified and compared with a standard value. If the amount of signal in the patient sample is significantly altered in comparison to a control sample then the presence of altered levels of nucleotide sequences encoding PKIN in the sample indicates the presence of the associated disorder. Such assays may also be used to evaluate the efficacy of a particular therapeutic treatment regimen in animal studies, in clinical trials, or to monitor the treatment of an individual patient.

[0258] In order to provide a basis for the diagnosis of a disorder associated with expression of PKIN, a normal or standard profile for expression is established. This may be accomplished by combining body fluids or cell extracts taken from normal subjects, either animal or human, with a sequence, or a fragment thereof, encoding PKIN, under conditions suitable for hybridization or amplification. Standard hybridization may be quantified by comparing the values obtained from normal subjects with values from an experiment in which a known amount of a substantially purified polynucleotide is used. Standard values obtained in this manner may be compared with values obtained from samples from patients who are symptomatic for a disorder. Deviation from standard values is used to establish the presence of a disorder.

[0259] Once the presence of a disorder is established and a treatment protocol is initiated, hybridization assays may be repeated on a regular basis to determine if the level of expression in the patient begins to approximate that which is observed in the normal subject. The results obtained from successive assays may be used to show the efficacy of treatment over a period ranging from several days to months.

[0260] With respect to cancer, the presence of an abnormal amount of transcript (either under- or overexpressed) in biopsied tissue from an individual may indicate a predisposition for the development of the disease, or may provide a means for detecting the disease prior to the appearance of actual clinical symptoms. A more definitive diagnosis of this type may allow health professionals to employ preventative measures or aggressive treatment earlier thereby preventing the development or further progression of the cancer.

[0261] Additional diagnostic uses for oligonucleotides designed from the sequences encoding PKIN may involve the use of PCR. These oligomers may be chemically synthesized, generated enzymatically, or produced in vitro. Oligomers will preferably contain a fragment of a polynucleotide encoding PKIN, or a fragment of a polynucleotide complementary to the polynucleotide encoding PKIN, and will be employed under optimized conditions for identification of a specific gene or condition. Oligomers may also be employed under less stringent conditions for detection or quantification of closely related DNA or RNA sequences.

[0262] In a particular aspect, oligonucleotide primers derived from the polynucleotide sequences encoding PKIN may be used to detect single nucleotide polymorphisms (SNPs). SNPs are substitutions, insertions and deletions that are a frequent cause of inherited or acquired genetic disease in humans. Methods of SNP detection include, but are not limited to, single-stranded conformation polymorphism (SSCP) and fluorescent SSCP (fSSCP) methods. In SSCP, oligonucleotide primers derived from the polynucleotide sequences encoding PKIN are used to amplify DNA using the polymerase chain reaction (PCR). The DNA may be derived, for example, from diseased or normal tissue, biopsy samples, bodily fluids, and the like. SNPs in the DNA cause differences in the secondary and tertiary structures of PCR products in single-stranded form, and these differences are detectable using gel electrophoresis in non-denaturing gels. In fSCCP, the oligonucleotide primers are fluorescently labeled, which allows detection of the amplimers in high-throughput equipment such as DNA sequencing machines. Additionally, sequence database analysis methods, termed in silico SNP (is SNP), are capable of identifying polymorphisms by comparing the sequence of individual overlapping DNA fragments which assemble into a common consensus sequence. These computer-based methods filter out sequence variations due to laboratory preparation of DNA and sequencing errors using statistical models and automated analyses of DNA sequence chromatograms. In the alternative, SNPs may be detected and characterized by mass spectrometry using, for example, the high throughput MASSARRAY system (Sequenom, Inc., San Diego Calif.).

[0263] Methods which may also be used to quantify the expression of PKIN include radiolabeling or biotinylating nucleotides, coamplification of a control nucleic acid, and interpolating results from standard curves. (See, e.g., Melby, P. C. et al. (1993) J. Immunol. Methods 159:235-244; Duplaa, C. et al. (1993) Anal. Biochem. 212:229-236.) The speed of quantitation of multiple samples may be accelerated by running the assay in a high-throughput format where the oligomer or polynucleotide of interest is presented in various dilutions and a spectrophotometric or colorimetric response gives rapid quantitation.

[0264] In further embodiments, oligonucleotides or longer fragments derived from any of the polynucleotide sequences described herein may be used as elements on a microarray. The microarray can be used in transcript imaging techniques which monitor the relative expression levels of large numbers of genes simultaneously as described below. The microarray may also be used to identify genetic variants, mutations, and polymorphisms. This information may be used to determine gene function, to understand the genetic basis of a disorder, to diagnose a disorder, to monitor progression/regression of disease as a function of gene expression, and to develop and monitor the activities of therapeutic agents in the treatment of disease. In particular, this information may be used to develop a pharmacogenomic profile of a patient in order to select the most appropriate and effective treatment regimen for that patient. For example, therapeutic agents which are highly effective and display the fewest side effects may be selected for a patient based on his/her pharmacogenomic profile.

[0265] In another embodiment, PKIN, fragments of PKIN, or antibodies specific for PKIN may be used as elements on a microarray. The microarray may be used to monitor or measure protein-protein interactions, drug-target interactions, and gene expression profiles, as described above.

[0266] A particular embodiment relates to the use of the polynucleotides of the present invention to generate a transcript image of a tissue or cell type. A transcript image represents the global pattern of gene expression by a particular tissue or cell type. Global gene expression patterns are analyzed by quantifying the number of expressed genes and their relative abundance under given conditions and at a given time. (See Seilhamer et al., "Comparative Gene Transcript Analysis," U.S. Pat. No. 5,840,484, expressly incorporated by reference herein.) Thus a transcript image may be generated by hybridizing the polynucleotides of the present invention or their complements to the totality of transcripts or reverse transcripts of a particular tissue or cell type. In one embodiment, the hybridization takes place in high-throughput format, wherein the polynucleotides of the present invention or their complements comprise a subset of a plurality of elements on a microarray. The resultant transcript image would provide a profile of gene activity.

[0267] Transcript images may be generated using transcripts isolated from tissues, cell lines, biopsies, or other biological samples. The transcript image may thus reflect gene expression in vivo, as in the case of a tissue or biopsy sample, or in vitro, as in the case of a cell line.

[0268] Transcript images which profile the expression of the polynucleotides of the present invention may also be used in conjunction with in vitro model systems and preclinical evaluation of pharmaceuticals, as well as toxicological testing of industrial and naturally-occurring environmental compounds. All compounds induce characteristic gene expression patterns, frequently termed molecular fingerprints or toxicant signatures, which are indicative of mechanisms of action and toxicity (Nuwaysir, E. F. et al. (1999) Mol. Carcinog. 24:153-159; Steiner, S. and N. L. Anderson (2000) Toxicol. Lett. 112-113:467-471, expressly incorporated by reference herein). If a test compound has a signature similar to that of a compound with known toxicity, it is likely to share those toxic properties. These fingerprints or signatures are most useful and refined when they contain expression information from a large number of genes and gene families. Ideally, a genome-wide measurement of expression provides the highest quality signature. Even genes whose expression is not altered by any tested compounds are important as well, as the levels of expression of these genes are used to normalize the rest of the expression data. The normalization procedure is useful for comparison of expression data after treatment with different compounds. While the assignment of gene function to elements of a toxicant signature aids in interpretation of toxicity mechanisms, knowledge of gene function is not necessary for the statistical matching of signatures which leads to prediction of toxicity. (See, for example, Press Release 00-02 from the National Institute of Environmental Health Sciences, released Feb. 29, 2000, available at http://www.niebs.nih.gov/oc/news/toxchip.htm.) Therefore, it is important and desirable in toxicological screening using toxicant signatures to include all expressed gene sequences.

[0269] In one embodiment, the toxicity of a test compound is assessed by treating a biological sample containing nucleic acids with the test compound. Nucleic acids that are expressed in the treated biological sample are hybridized with one or more probes specific to the polynucleotides of the present invention, so that transcript levels corresponding to the polynucleotides of the present invention may be quantified. The transcript levels in the treated biological sample are compared with levels in an untreated biological sample. Differences in the transcript levels between the two samples are indicative of a toxic response caused by the test compound in the treated sample.

[0270] Another particular embodiment relates to the use of the polypeptide sequences of the present invention to analyze the proteome of a tissue or cell type. The term proteome refers to the global pattern of protein expression in a particular tissue or cell type. Each protein component of a proteome can be subjected individually to further analysis. Proteome expression patterns, or profiles, are analyzed by quantifying the number of expressed proteins and their relative abundance under given conditions and at a given time. A profile of a cell's proteome may thus be generated by separating and analyzing the polypeptides of a particular tissue or cell type. In one embodiment, the separation is achieved using two-dimensional gel electrophoresis, in which proteins from a sample are separated by isoelectric focusing in the first dimension, and then according to molecular weight by sodium dodecyl sulfate slab gel electrophoresis in the second dimension (Steiner and Anderson, supra). The proteins are visualized in the gel as discrete and uniquely positioned spots, typically by staining the gel with an agent such as Coomassie Blue or silver or fluorescent stains. The optical density of each protein spot is generally proportional to the level of the protein in the sample. The optical densities of equivalently positioned protein spots from different samples, for example, from biological samples either treated or untreated with a test compound or therapeutic agent, are compared to identify any changes in protein spot density related to the treatment. The proteins in the spots are partially sequenced using, for example, standard methods employing chemical or enzymatic cleavage followed by mass spectrometry. The identity of the protein in a spot may be determined by comparing its partial sequence, preferably of at least 5 contiguous amino acid residues, to the polypeptide sequences of the present invention. In some cases, further sequence data may be obtained for definitive protein identification.

[0271] A proteomic profile may also be generated using antibodies specific for PKIN to quantify the levels of PKIN expression. In one embodiment, the antibodies are used as elements on a microarray, and protein expression levels are quantified by exposing the microarray to the sample and detecting the levels of protein bound to each array element (Lueking, A. et al. (1999) Anal. Biochem. 270:103-111; Mendoze, L. G. et al. (1999) Biotechniques 27:778-788). Detection may be performed by a variety of methods known in the art, for example, by reacting the proteins in the sample with a thiol- or amino-reactive fluorescent compound and detecting the amount of fluorescence bound at each array element.

[0272] Toxicant signatures at the proteome level are also useful for toxicological screening, and should be analyzed in parallel with toxicant signatures at the transcript level. There is a poor correlation between transcript and protein abundances for some proteins in some tissues (Anderson, N. J. and J. Seilhamner (1997) Electrophoresis 18:533-537), so proteome toxicant signatures may be useful in the analysis of compounds which do not significantly affect the transcript image, but which alter the proteomic profile. In addition, the analysis of transcripts in body fluids is difficult, due to rapid degradation of mRNA, so proteomic profiling may be more reliable and informative in such cases.

[0273] In another embodiment, the toxicity of a test compound is assessed by treating a biological sample containing proteins with the test compound. Proteins that are expressed in the treated biological sample are separated so that the amount of each protein can be quantified. The amount of each protein is compared to the amount of the corresponding protein in an untreated biological sample. A difference in the amount of protein between the two samples is indicative of a toxic response to the test compound in the treated sample. Individual proteins are identified by sequencing the amino acid residues of the individual proteins and comparing these partial sequences to the polypeptides of the present invention.

[0274] In another embodiment, the toxicity of a test compound is assessed by treating a biological sample containing proteins with the test compound. Proteins from the biological sample are incubated with antibodies specific to the polypeptides of the present invention. The amount of protein recognized by the antibodies is quantified. The amount of protein in the treated biological sample is compared with the amount in an untreated biological sample. A difference in the amount of protein between the two samples is indicative of a toxic response to the test compound in the treated sample.

[0275] Microarrays may be prepared, used, and analyzed using methods known in the art. (See, e.g., Brennan, T. M. et al. (1995) U.S. Pat. No. 5,474,796; Schena, M. et al. (1996) Proc. Natl. Acad. Sci. USA 93:10614-10619; Baldeschweiler et al. (1995) PCT application WO95/251116; Shalon, D. et al. (1995) PCT application WO95/35505; Heller, R. A. et al. (1997) Proc. Natl. Acad. Sci. USA 94:2150-2155; and Heller, M. J. et al. (1997) U.S. Pat. No. 5,605,662.) Various types of microarrays are well known and thoroughly described in DNA Microarrays: A Practical Approach, M. Schena, ed. (1999) Oxford University Press, London, hereby expressly incorporated by reference.

[0276] In another embodiment of the invention, nucleic acid sequences encoding PKIN may be used to generate hybridization probes useful in mapping the naturally occurring genomic sequence. Either coding or noncoding sequences may be used, and in some instances, noncoding sequences may be preferable over coding sequences. For example, conservation of a coding sequence among members of a multi-gene family may potentially cause undesired cross hybridization during chromosomal mapping. The sequences may be mapped to a particular chromosome, to a specific region of a chromosome, or to artificial chromosome constructions, e.g., human artificial chromosomes (HACs), yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), bacterial P1 constructions, or single chromosome cDNA libraries. (See, e.g., Harrington, J. J. et al. (1997) Nat. Genet. 15:345-355; Price, C. M. (1993) Blood Rev. 7:127-134; and Trask, B. J. (1991) Trends Genet. 7:149-154.) Once mapped, the nucleic acid sequences of the invention may be used to develop genetic linkage maps, for example, which correlate the inheritance of a disease state with the inheritance of a particular chromosome region or restriction fragment length polymorphism (RFLP). (See, for example, Lander, E. S. and D. Botstein (1986) Proc. Natl. Acad. Sci. USA 83:7353-7357.) Fluorescent in situ hybridization (FISH) may be correlated with other physical and genetic map data. (See, e.g., Heinz-Ulrich, et al. (1995) in Meyers, supra, pp. 965-968.) Examples of genetic map data can be found in various scientific journals or at the Online Mendelian Inheritance in Man (OMIM) World Wide Web site. Correlation between the location of the gene encoding PKIN on a physical map and a specific disorder, or a predisposition to a specific disorder, may help define the region of DNA associated with that disorder and thus may further positional cloning efforts.

[0277] In situ hybridization of chromosomal preparations and physical mapping techniques, such as linkage analysis using established chromosomal markers, may be used for extending genetic maps. Often the placement of a gene on the chromosome of another mammalian species, such as mouse, may reveal associated markers even if the exact chromosomal locus is not known. This information is valuable to investigators searching for disease genes using positional cloning or other gene discovery techniques. Once the gene or genes responsible for a disease or syndrome have been crudely localized by genetic linkage to a particular genomic region, e.g., ataxia-telangiectasia to 11q22-23, any sequences mapping to that area may represent associated or regulatory genes for further investigation. (See, e.g., Gatti, R. A. et al. (1988) Nature 336:577-580.) The nucleotide sequence of the instant invention may also be used to detect differences in the chromosomal location due to translocation, inversion, etc., among normal, carrier, or affected individuals.

[0278] In another embodiment of the invention, PKIN, its catalytic or immunogenic fragments, or oligopeptides thereof can be used for screening libraries of compounds in any of a variety of drug screening techniques. The fragment employed in such screening may be free in solution, affixed to a solid support, borne on a cell surface, or located intracellularly. The formation of binding complexes between PKIN and the agent being tested may be measured.

[0279] Another technique for drug screening provides for high throughput screening of compounds having suitable binding affinity to the protein of interest. (See, e.g., Geysen, et al. (1984) PCT application WO84/03564.) In this method, large numbers of different small test compounds are synthesized on a solid substrate. The test compounds are reacted with PKIN, or fragments thereof, and washed. Bound PKIN is then detected by methods well known in the art. Purified PKIN can also be coated directly onto plates for use in the aforementioned drug screening techniques. Alternatively, non-neutralizing antibodies can be used to capture the peptide and immobilize it on a solid support.

[0280] In another embodiment, one may use competitive drug screening assays in which neutralizing antibodies capable of binding PKIN specifically compete with a test compound for binding PKIN. In this manner, antibodies can be used to detect the presence of any peptide which shares one or more antigenic determinants with PKIN.

[0281] In additional embodiments, the nucleotide sequences which encode PKIN may be used in any molecular biology techniques that have yet to be developed, provided the new techniques rely on properties of nucleotide sequences that are currently known, including, but not limited to, such properties as the triplet genetic code and specific base pair interactions.

[0282] Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. The following embodiments are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.

[0283] Without further elaboration, it is believed that one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. The following preferred specific embodiments are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever.

[0284] The disclosures of all patents, applications, and publications mentioned above and below, including U.S. Ser. No. 60/199,021, U.S. Ser. No. 60/200,226, U.S. Ser. No. 60/202,339, U.S. Ser. No. 60/203,505, U.S. Ser. No. 60/205,654, U.S. Ser. No. 60/207,739, and U.S. Ser. No. 60/208,795, are hereby expressly incorporated by reference.

EXAMPLES

[0285] 1. Construction of cDNA Libraries

[0286] Incyte cDNAs were derived from cDNA libraries described in the LIFESEQ GOLD database (Incyte Genomics, Palo Alto Calif.) and shown in Table 4, column 5. Some tissues were homogenized and lysed in guanidinium isothiocyanate, while others were homogenized and lysed in phenol or in a suitable mixture of denaturants, such as TRIZOL (Life Technologies), a monophasic solution of phenol and guanidine isothiocyanate. The resulting lysates were centrifuged over CsCl cushions or extracted with chloroform. RNA was precipitated from the lysates with either isopropanol or sodium acetate and ethanol, or by other routine methods.

[0287] Phenol extraction and precipitation of RNA were repeated as necessary to increase RNA purity. In some cases, RNA was treated with DNase. For most libraries, poly(A).sup.+ RNA was isolated using oligo d(T)coupled paramagnetic particles (Promega), OLIGOTEX latex particles (QIAGEN, Chatsworth Calif.), or an OLIGOTEX mRNA purification kit (QIAGEN). Alternatively, RNA was isolated directly from tissue lysates using other RNA isolation kits, e.g., the POLY(A)PURE mRNA purification kit (Ambion, Austin Tex.). in some cases, Stratagene was provided with RNA and constructed the corresponding cDNA libraries. Otherwise, cDNA was synthesized and cDNA libraries were constructed with the UNIAP vector system (Stratagene) or SUPERSCRIPT plasmid system (Life Technologies), using the recommended procedures or similar methods known in the art. (See, e.g., Ausubel, 1997, supra, units 5.10.6.) Reverse transcription was initiated using oligo d(T) or random primers. Synthetic oligonucleotide adapters were ligated to double stranded cDNA, and the cDNA was digested with the appropriate restriction enzyme or enzymes. For most libraries, the cDNA was size-selected (300-1000 bp) using SEPHACRYL S1000, SEPHAROSE CL2B, or SEPHAROSE CL4B column chromatography (Amersham Pharmacia Biotech) or preparative agarose gel electrophoresis. cDNAs were ligated into compatible restriction enzyme sites of the polylinker of a suitable plasmid, e.g., PBLUESCRIPT plasmid (Stratagene), PSPORT1 plasmid (Life Technologies), PCDNA2.1 plasmid (Invitrogen, Carlsbad Calif.), PBK-CMV plasmid (Stratagene), or pINCY (Incyte Genomics, Palo Alto Calif.), or derivatives thereof Recombinant plasmids were transformed into competent E. coli cells including XL1-Blue, XL1-BlueMRF, or SOLR from Stratagene or DHS.alpha., DH10B, or ElectroMAX DH10B from Life Technologies.

[0288] II. Isolation of cDNA Clones

[0289] Plasmids obtained as described in Example I were recovered from host cells by in vivo excision using the UNIAP vector system (Stratagene) or by cell lysis. Plasmids were purified using at least one of the following: a Magic or WIZARD Minipreps DNA purification system (Promega); an AGTC Miniprep purification kit (Edge Biosystems, Gaithersburg Md.); and QIAWELL 8 Plasmid, QIAWELL 8 Plus Plasmid, QIAWELL 8 Ultra Plasmid purification systems or the R.E.A.L. PREP 96 plasmid purification kit from QIAGEN. Following precipitation, plasmids were resuspended in 0.1 ml of distilled water and stored, with or without lyophilization, at 4.degree. C.

[0290] Alternatively, plasmid DNA was amplified from host cell lysates using direct link PCR in a high-throughput format (Rao, V. B. (1994) Anal. Biochem. 216:1-14). Host cell lysis and thermal cycling steps were carried out in a single reaction mixture Samples were processed and stored in 384-well plates, and the concentration of amplified plasmid DNA was quantified fluorometrically using PICOGREEN dye (Molecular Probes, Eugene Oreg.) and a FLUOROSKAN II fluorescence scanner (Labsystems Oy, Helsinki, Finland).

[0291] III. Sequencing and Analysis

[0292] Incyte cDNA recovered in plasmids as described in Example II were sequenced as follows. Sequencing reactions were processed using standard methods or high-throughput instrumentation such as the ABI CATALYST 800 (Applied Biosystems) thermal cycler or the PTC-200 thermal cycler (MJ Research) in conjunction with the HYDRA microdispenser (Robbins Scientific) or the MICROLAB 2200 (Hamilton) liquid transfer system. cDNA sequencing reactions were prepared using reagents provided by Amersham Pharmacia Biotech or supplied in ABI sequencing kits such as the ABI PRISM BIGDYE Terminator cycle sequencing ready reaction kit (Applied Biosystems). Electrophoretic separation of cDNA sequencing reactions and detection of labeled polynucleotides were carried out using the MEGABACE 1000 DNA sequencing system (Molecular Dynamics); the ABI PRISM 373 or 377 sequencing system (Applied Biosystems) in conjunction with standard ABI protocols and base calling software; or other sequence analysis systems known in the art. Reading frames within the cDNA sequences were identified using standard methods (reviewed in Ausubel, 1997, supra, unit 7.7). Some of the cDNA sequences were selected for extension using the techniques disclosed in Example VIII.

[0293] The polynucleotide sequences derived from Incyte cDNAs were validated by removing vector, linker, and poly(A) sequences and by masking ambiguous bases, using algorithms and programs based on BLAST, dynamic programming, and dinucleotide nearest neighbor analysis. The Incyte cDNA sequences or translations thereof were then queried against a selection of public databases such as the GenBank primate, rodent, mammalian, vertebrate, and eukaryote databases, and BLOCKS, PRINTS, DOMO, PRODOM, and hidden Markov model (HMM)-based protein family databases such as PFAM. (HMM is a probabilistic approach which analyzes consensus primary structures of gene families. See, for example, Eddy, S. R. (1996) Curr. Opin. Struct. Biol. 6:361-365.) The queries were performed using programs based on BLAST, FASTA, BLIMPS, and HMMER. The Incyte cDNA sequences were assembled to produce full length polynucleotide sequences. Alternatively, GenBank cDNAs, GenBank ESTs, stitched sequences, stretched sequences, or Genscan-predicted coding sequences (see Examples IV and V) were used to extend Incyte cDNA assemblages to full length. Assembly was performed using programs based on Phred, Phrap, and Consed, and cDNA assemblages were screened for open reading frames using programs based on GeneMark, BLAST, and FASTA. The full length polynucleotide sequences were translated to derive the corresponding full length polypeptide sequences. Alternatively, a polypeptide of the invention may begin at any of the methionine residues of the full length translated polypeptide. Full length polypeptide sequences were subsequently analyzed by querying against databases such as the GenBank protein databases (genpept), SwissProt, BLOCKS, PRINTS, DOMO, PRODOM, Prosite, and hidden Markov model (HMM)-based protein family databases such as PPAM. Full length polynucleotide sequences are also analyzed using MACDNASIS PRO software (Hitachi Software Engineering, South San Francisco Calif.) and LASERGENE software (DNASTAR). Polynucleotide and polypeptide sequence alignments are generated using default parameters specified by the CLUSTAL algorithm as incorporated into the MEGALIGN multisequence alignment program (DNASTAR), which also calculates the percent identity between aligned sequences.

[0294] Table 7 summarizes the tools, programs, and algorithms used for the analysis and assembly of Incyte cDNA and full length sequences and provides applicable descriptions, references, and threshold parameters. The first column of Table 7 shows the tools, programs, and algorithms used, the second column provides brief descriptions thereof, the third column presents appropriate references, all of which are incorporated by reference herein in their entirety, and the fourth column presents, where applicable, the scores, probability values, and other parameters used to evaluate the strength of a match between two sequences (the higher the score or the lower the probability value, the greater the identity between two sequences).

[0295] The programs described above for the assembly and analysis of full length polynucleotide and polypeptide sequences were also used to identify polynucleotide sequence fragments from SEQ ID NO:19-36. Fragments from about 20 to about 4000 nucleotides which are useful in hybridization and amplification technologies are described in Table 4, column 4.

[0296] IV. Identification and Editing of Coding Sequences from Genomic DNA

[0297] Putative human kinases were initially identified by running the Genscan gene identification program against public genomic sequence databases (e.g., gbpri and gbhtg). Genscan is a general-purpose gene identification program which analyzes genomic DNA sequences from a variety of organisms (See Burge, C. and S. Karlin (1997) J. Mol. Biol. 268:78-94, and Burge, C. and S. Karlin (1998) Curr. Opin. Struct. Biol. 8:346354). The program concatenates predicted exons to form an assembled cDNA sequence extending from a methionine to a stop codon. The output of Genscan is a FASTA database of polynucleotide and polypeptide sequences. The maximum range of sequence for Genscan to analyze at once was set to 30 kb. To determine which of these Genscan predicted cDNA sequences encode human kinases, the encoded polypeptides were analyzed by querying against PFAM models for kinases. Potential human kinases were also identified by homology to Incyte cDNA sequences that had been annotated as kinases. These selected Genscan-predicted sequences were then compared by BLAST analysis to the genpept and gbpri public databases. Where necessary, the Genscan-predicted sequences were then edited by comparison to the top BLAST hit from genpept to correct errors in the sequence predicted by Genscan, such as extra or omitted exons. BLAST analysis was also used to find any Incyte cDNA or public cDNA coverage of the Genscan-predicted sequences, thus providing evidence for transcription. When Incyte cDNA coverage was available, this information was used to correct or confirm the Genscan predicted sequence. Full length polynucleotide sequences were obtained by assembling Genscan-predicted coding sequences with Incyte cDNA sequences and/or public cDNA sequences using the assembly process described in Example III. Alternatively, full length polynucleotide sequences were derived entirely from edited or unedited Genscan-predicted coding sequences.

[0298] V. Assembly of Genomic Sequence Data with cDNA Sequence Data

[0299] "Stitched" Sequences

[0300] Partial cDNA sequences were extended with exons predicted by the Genscan gene identification program described in Example IV. Partial cDNAs assembled as described in Example m were mapped to genomic DNA and parsed into clusters containing related cDNAs and Genscan exon predictions from one or more genomic sequences. Each cluster was analyzed using an algorithm based on graph theory and dynamic programming to integrate cDNA and genomic information, generating possible splice variants that were subsequently confirmed, edited, or extended to create a full length sequence. Sequence intervals in which the entire length of the interval was present on more than one sequence in the cluster were identified, and intervals thus identified were considered to be equivalent by transitivity. For example, if an interval was present on a cDNA and two genomic sequences, then all three intervals were considered to be equivalent. This process allows unrelated but consecutive genomic sequences to be brought together, bridged by cDNA sequence. Intervals thus identified were then "stitched" together by the stitching algorithm in the order that they appear along their parent sequences to generate the longest possible sequence, as well as sequence variants. Linkages between intervals which proceed along one type of parent sequence (cDNA to cDNA or genomic sequence to genomic sequence) were given preference over linkages which change parent type (cDNA to genomic sequence). The resultant stitched sequences were translated and compared by BLAST analysis to the genpept and gbpri public databases. Incorrect exons predicted by Genscan were corrected by comparison to the top BLAST hit from genpept. Sequences were further extended with additional cDNA sequences, or by inspection of genomic DNA, when necessary.

[0301] "Stretched" Sequences

[0302] Partial DNA sequences were extended to full length with an algorithm based on BLAST analysis. First, partial cDNAs assembled as described in Example III were queried against public databases such as the GenBank primate, rodent, mammalian, vertebrate, and eukaryote databases using the BLAST program. The nearest GenBank protein homolog was then compared by BLAST analysis to either Incyte cDNA sequences or GenScan exon predicted sequences described in Example IV. A chimeric protein was generated by using the resultant high-scoring segment pairs (HSPs) to map the translated sequences onto the GenBank protein homolog. Insertions or deletions may occur in the chimeric protein with respect to the original GenBank protein homolog. The GenBank protein homolog, the chimeric protein, or both were used as probes to search for homologous genomic sequences from the public human genome databases. Partial DNA sequences were therefore "stretched" or extended by the addition of homologous genomic sequences. The resultant stretched sequences were examined to determine whether it contained a complete gene.

[0303] VI. Chromosomal Mapping of PKIN Encoding Polynucleotides

[0304] The sequences which were used to assemble SEQ ID NO:19-36 were compared with sequences from the Incyte LIFESEQ database and public domain databases using BLAST and other implementations of the Smith-Waterman algorithm. Sequences from these databases that matched SEQ ID NO:19-36 were assembled into clusters of contiguous and overlapping sequences using assembly algorithms such as Phrap (Table 7). Radiation hybrid and genetic mapping data available from public resources such as the Stanford Human Genome Center (SHGC), Whitehead Institute for Genome Research (WIGR), and Genethon were used to determine if any of the clustered sequences had been previously mapped. Inclusion of a mapped sequence in a cluster resulted in the assignment of all sequences of that cluster, including its particular SEQ ID NO:, to that map location.

[0305] Map locations are represented by ranges, or intervals, of human chromosomes. The map position of an interval, in centiMorgans, is measured relative to the terminus of the chromosome's p-arm. (The centiMorgan (cM) is a unit of measurement based on recombination frequencies between chromosomal markers. On average, 1 cM is roughly equivalent to 1 megabase (Mb) of DNA in humans, although this can vary widely due to hot and cold spots of recombination.) The cM distances are based on genetic markers mapped by Genethon which provide boundaries for radiation hybrid markers whose sequences were included in each of the clusters. Human genome maps and other resources available to the public, such as the NCBI "GeneMap'99" World Wide Web site (http://www.ncbi.nlm.ni- h.gov/genemap/), can be employed to determine if previously identified disease genes map within or in proximity to the intervals indicated above.

[0306] In this manner, SEQ ID NO:24 was mapped to chromosome 2 within the interval from 92.30 to 103.1 centiMorgans, SEQ ID NO:25 was mapped to chromosome 11 within the interval from 104.8 to 117.9 centiMorgans, SEQ ID NO:33 was mapped to chromosome 8 within the interval from 25.8 to 40.3 centiMorgans, SEQ ID NO:23 was mapped to chromosome 9 within the interval from 101.20 to 104.90 centiMorgans, to chromosome 10 within the interval from 145.20 to 156.60 centiMorgans, and to chromosome 19 within the interval from 69.90 to 81.20 centiMorgans. More than one map location is reported for SEQ ID NO:23, indicating that sequences having different map locations were assembled into a single cluster. This situation occurs when sequences having strong similarity, but not complete identity, are assembled into a single cluster.

[0307] VII. Analysis of Polynucleotide Expression

[0308] Northern analysis is a laboratory technique used to detect the presence of a transcript of a gene and involves the hybridization of a labeled nucleotide sequence to a membrane on which RNAs from a particular cell type or tissue have been bound. (See, e.g., Sambrook, supra, ch. 7; Ausubel (1995) supra, ch. 4 and 16.)

[0309] Analogous computer techniques applying BLAST were used to search for identical or related molecules in cDNA databases such as GenBank or LIFESEQ (Incyte Genomics). This analysis is much faster than multiple membrane-based hybridizations. In addition, the sensitivity of the computer search can be modified to determine whether any particular match is categorized as exact or similar. The basis of the search is the product score, which is defined as: 1 BLAST Score .times. Percent Identity 5 .times. minimum { length ( Seq . 1 ) , length ( Seq . 2 ) }

[0310] The product score takes into account both the degree of similarity between two sequences and the length of the sequence match. The product score is a normalized value between 0 and 100, and is calculated as follows: the BLAST score is multiplied by the percent nucleotide identity and the product is divided by (5 times the length of the shorter of the two sequences). The BLAST score is calculated by assigning a score of +5 for every base that matches in a high-scoring segment pair (HSP), and 4 for every mismatch. Two sequences may share more than one HSP (separated by gaps). If there is more than one HSP, then the pair with the highest BLAST score is used to calculate the product score. The product score represents a balance between fractional overlap and quality in a BLAST alignment. For example, a product score of 100 is produced only for 100% identity over the entire length of the shorter of the two sequences being compared. A product score of 70 is produced either by 100% identity and 70% overlap at one end, or by 88% identity and 100% overlap at the other. A product score of 50 is produced either by 100% identity and 50% overlap at one end, or 79% identity and 100% overlap.

[0311] Alternatively, polynucleotide sequences encoding PKIN are analyzed with respect to the tissue sources from which they were derived. For example, some full length sequences are assembled, at least in part, with overlapping Incyte cDNA sequences (see Example III). Each cDNA sequence is derived from a cDNA library constructed from a human tissue. Each human tissue is classified into one of the following organ/tissue categories: cardiovascular system; connective tissue; digestive system; embryonic structures; endocrine system; exocrine glands; genitalia, female; genitalia, male; germ cells; hemic and immune system; liver; musculoskeletal system; nervous system; pancreas; respiratory system; sense organs; skin; stomatognathic system; unclassified/mixed; or urinary tract. The number of libraries in each category is counted and divided by the total number of libraries across all categories. Similarly, each human tissue is classified into one of the following disease/condition categories: cancer, cell line, developmental, inflammation, neurological, trauma, cardiovascular, pooled, and other, and the number of libraries in each category is counted and divided by the total number of libraries across all categories. The resulting percentages reflect the tissue- and disease-specific expression of cDNA encoding PKIN. cDNA sequences and cDNA library/tissue information are found in the LIFESEQ GOLD database (Incyte Genomics, Palo Alto Calif.).

[0312] VIII. Extension of PKIN Encoding Polynucleotides

[0313] Pull length polynucleotide sequences were also produced by extension of an appropriate fragment of the full length molecule using oligonucleotide primers designed from this fragment. One primer was synthesized to initiate 5' extension of the known fragment, and the other primer was synthesized to initiate 3' extension of the known fragment. The initial primers were designed using OLIGO 4.06 software (National Biosciences), or another appropriate program, to be about 22 to 30 nucleotides in length, to have a GC content of about 50% or more, and to anneal to the target sequence at temperatures of about 68.degree. C. to about 72.degree. C. Any stretch of nucleotides which would result in hairpin structures and primer-primer dimerizations was avoided.

[0314] Selected human cDNA libraries were used to extend the sequence. If more than one extension was necessary or desired, additional or nested sets of primers were designed.

[0315] High fidelity amplification was obtained by PCR using methods well known in the art. PCR was performed in 96-well plates using the PTC-200 thermal cycler (MJ Research, Inc.). The reaction mix contained DNA template, 200 mmol of each primer, reaction buffer containing Me.sup.2+, (NH.sub.4).sub.2SO.sub.4, and 2-mercaptoethanol, Taq DNA polymerase (Amersham Pharmacia Biotech), ELONGASE enzyme (Life Technologies), and Pfu DNA polymerase (Stratagene), with the following parameters for primer pair PCI A and PCI B: Step 1: 94.degree. C., 3 min; Step 2: 94.degree. C., 15 sec; Step 3: 60.degree. C., 1 min; Step 4: 68.degree. C., 2 min; Step 5: Steps 2, 3, and 4 repeated 20 times; Step 6: 68.degree. C., 5 min; Step 7: storage at 4.degree. C. In the alternative, the parameters for primer pair T7 and SK+ were as follows: Step 1: 94.degree. C., 3 min; Step 2: 94.degree. C., 15 sec; Step 3: 57.degree. C., 1 min; Step 4: 68.degree. C., 2 min; Step 5: Steps 2, 3, and 4 repeated 20 times;

[0316] Step 6: 68.degree. C., 5 min; Step 7: storage at 4.degree. C.

[0317] The concentration of DNA in each well was determined by dispensing 100 .mu.l PICOGREEN quantitation reagent (0.25% (v/v) PICOGREEN; Molecular Probes, Eugene Oreg.) dissolved in 1.times.TE and 0.5 .mu.l of undiluted PCR product into each well of an opaque fluorimeter plate (Corning Costar, Acton Mass.), allowing the DNA to bind to the reagent. The plate was scanned in a Fluoroskan II (Labsystems Oy, Helsinki, Finland) to measure the fluorescence of the sample and to quantify the concentration of DNA. A 5 .mu.l to 10 .mu.l aliquot of the reaction mixture was analyzed by electrophoresis on a 1% agarose gel to determine which reactions were successful in extending the sequence.

[0318] The extended nucleotides were desalted and concentrated, transferred to 384-well plates, digested with CviJI cholera virus endonuclease (Molecular Biology Research, Madison Wis.), and sonicated or sheared prior to religation into pUC 18 vector (Amersham Pharmacia Biotech). For shotgun sequencing, the digested nucleotides were separated on low concentration (0.6 to 0.8%) agarose gels, fragments were excised, and agar digested with Agar ACE (Promega). Extended clones were religated using T4 ligase (New England Biolabs, Beverly Mass.) into pUC 18 vector (Amersham Pharmacia Biotech), treated with Pfu DNA polymerase (Stratagene) to fill-in restriction site overhangs, and transfected into competent E. coli cells. Transformed cells were selected on antibiotic-containing media, and individual colonies were picked and cultured overnight at 37.degree. C. in 384-well plates in LB/2.times. carb liquid media.

[0319] The cells were lysed, and DNA was amplified by PCR using Taq DNA polymerase (Amersham Pharmacia Biotech) and Pfu DNA polymerase (Stratagene) with the following parameters: Step 1: 94.degree. C., 3 min; Step 2: 94.degree. C., 15 sec; Step 3: 60.degree. C., 1 min; Step 4: 72.degree. C., 2 min;

[0320] Step 5: steps 2, 3, and 4 repeated 29 times; Step 6: 72.degree. C., 5 min; Step 7: storage at 4.degree. C. DNA was quantified by PICOGREEN reagent (Molecular Probes) as described above. Samples with low DNA recoveries were reamplified using the same conditions as described above. Samples were diluted with 20% dimethysulfoxide (1:2, v/v), and sequenced using DYENAMIC energy transfer sequencing primers and the DYENAMIC DIRECT kit (Amersham Pharmacia Biotech) or the ABI PRISM BIGDYE Terminator cycle sequencing ready reaction kit (Applied Biosystems).

[0321] In like manner, full length polynucleotide sequences are verified using the above procedure or are used to obtain 5' regulatory sequences using the above procedure along with oligonucleotides designed for such extension, and an appropriate genomic library.

[0322] IK. Labeling and Use of Individual Hybridization Probes

[0323] Hybridization probes derived from SEQ ID NO:19-36 are employed to screen cDNAs, genomic DNAs, or mRNAs. Although the labeling of oligonucleotides, consisting of about 20 base pairs, is specifically described, essentially the same procedure is used with larger nucleotide fragments. Oligonucleotides are designed using state-of-the-art software such as OLIGO 4.06 software (National Biosciences) and labeled by combining 50 pmol of each oligomer, 250 .mu.Ci of [.gamma.-.sup.32P] adenosine triphosphate (Amersham Pharmacia Biotech), and T4 polynucleotide kinase (DuPont NEN, Boston Mass.). The labeled oligonucleotides are substantially purified using a SEPHADEX G-25 superfine size exclusion dextran bead column (Amersham Pharmacia Biotech). An aliquot containing 10.sup.7 counts per minute of the labeled probe is used in a typical membrane-based hybridization analysis of human genomic DNA digested with one of the following endonucleases: Ase I, Bgl II, Eco RI, Pst I, Xba I, or Pvu II (DuPont NEN).

[0324] The DNA from each digest is fractionated on a 0.7% agarose gel and transferred to nylon membranes (Nytran Plus, Schleicher & Schuell, Durham N.H.). Hybridization is carried out for 16 hours at 40.degree. C. To remove nonspecific signals, blots are sequentially washed at room temperature under conditions of up to, for example, 0.1.times.saline sodium citrate and 0.5% sodium dodecyl sulfate. Hybridization patterns are visualized using autoradiography or an alternative imaging means and compared.

[0325] X. Microarrays

[0326] The linkage or synthesis of array elements upon a microarray can be achieved utilizing photolithography, piezoelectric printing (ink-jet printing, See, e.g., Baldeschweiler, supra.), mechanical microspotting technologies, and derivatives thereof. The substrate in each of the aforementioned technologies should be uniform and solid with a non-porous surface (Schena (1999), supra). Suggested substrates include silicon, silica, glass slides, glass chips, and silicon wafers. Alternatively, a procedure analogous to a dot or slot blot may also be used to arrange and link elements to the surface of a substrate using thermal, UV, chemical, or mechanical bonding procedures. A typical array may be produced using available methods and machines well known to those of ordinary skill in the art and may contain any appropriate number of elements. (See, e.g., Schena, M. et al. (1995) Science 270:467-470; Shalon, D. et al. (1996) Genome Res. 6:639-645; Marshall, A. and S. Hodgson (1998) Nat. Biotechnol. 16:27-31.) Full length cDNAs, Expressed Sequence Tags (ESTs), or fragments or oligomers thereof may comprise the elements of the microarray. Fragments or oligomers suitable for hybridization can be selected using software well known in the art such as LASERGENE software (DNASTAR). The array elements are hybridized with polynucleotides in a biological sample. The polynucleotides in the biological sample are conjugated to a fluorescent label or other molecular tag for ease of detection. After hybridization, nonhybridized nucleotides from the biological sample are removed, and a fluorescence scanner is used to detect hybridization at each array element. Alternatively, laser desorbtion and mass spectrometry may be used for detection of hybridization. The degree of complementarity and the relative abundance of each polynucleotide which hybridizes to an element on the microarray may be assessed. In one embodiment, microarray preparation and usage is described in detail below.

[0327] Tissue or Cell Sample Preparation

[0328] Total RNA is isolated from tissue samples using the guanidinium thiocyanate method and poly(A).sup.+ RNA is purified using the oligo-(dT) cellulose method. Each poly(A).sup.+ RNA sample is reverse transcribed using MMLV reverse-transcriptase, 0.05 .mu.g/.mu.l oligo(dT) primer (21mer), 1.times. first strand buffer, 0.03 units/.mu.l RNase inhibitor, 500 .mu.M dATP, 500 .mu.M dGTP, 500 .mu.M dTTP, 40 .mu.M dCTP, 40 .mu.M dCTP-Cy3 (BDS) or dCTP-Cy5 (Amersham Pharmacia Biotech). The reverse transcription reaction is performed in a 25 ml volume containing 200 ng poly(A).sup.+ RNA with GEMBRIGHT kits (Incyte). Specific control poly(A).sup.+ RNAs are synthesized by in vitro transcription from non-coding yeast genomic DNA. After incubation at 37.degree. C. for 2 hr, each reaction sample (one with Cy3 and another with Cy5 labeling) is treated with 2.5 ml of 0.5M sodium hydroxide and incubated for 20 minutes at 85.degree. C. to the stop the reaction and degrade the RNA. Samples are purified using two successive CHROMA SPIN 30 gel filtration spin columns (CLONIECH Laboratories, Inc. (CLONTECH), Palo Alto Calif.) and after combining, both reaction samples are ethanol precipitated using 1 ml of glycogen (1 mg/ml), 60 ml sodium acetate, and 300 ml of 100% ethanol. The sample is then dried to completion using a SpeedVAC (Savant Instruments Inc., Holbrook N.Y.) and resuspended in 14 pi 5.times.SSC/0.2% SDS.

[0329] Microarray Preparation

[0330] Sequences of the present invention are used to generate array elements. Each array element is amplified from bacterial cells containing vectors with cloned cDNA inserts. PCR amplification uses primers complementary to the vector sequences flanking the cDNA insert. Array elements are amplified in thirty cycles of PCR from an initial quantity of 1-2 ng to a final quantity greater than 5 Lg. Amplified array elements are then purified using SEPHACRYL-400 (Amersham Pharmacia Biotech).

[0331] Purified array elements are immobilized on polymer-coated glass slides. Glass microscope slides (Corning) are cleaned by ultrasound in 0.1% SDS and acetone, with extensive distilled water washes between and after treatments. Glass slides are etched in 4% hydrofluoric acid (VWR Scientific Products Corporation (VWR), West Chester Pa.), washed extensively in distilled water, and coated with 0.05% aminopropyl silane (Sigma) in 95% ethanol. Coated slides are cured in a 110.degree. C. oven.

[0332] Array elements are applied to the coated glass substrate using a procedure described in U.S. Pat. No. 5,807,522, incorporated herein by reference. 1 .mu.l of the array element DNA, at an average concentration of 100 ng/.mu.l, is loaded into the open capillary printing element by a high-speed robotic apparatus. The apparatus then deposits about 5 nl of array element sample per slide.

[0333] Microarrays are UV-crosslinked using a STRATALINER UV-crosslinker (Stratagene). Microarrays are washed at room temperature once in 0.2% SDS and three times in distilled water. Non-specific binding sites are blocked by incubation of microarrays in 0.2% casein in phosphate buffered saline (PBS) (Tropix, Inc., Bedford Mass.) for 30 minutes at 60.degree. C. followed by washes in 0.2% SDS and distilled water as before.

[0334] Hybridization

[0335] Hybridization reactions contain 9 .mu.l of sample mixture consisting of 0.2 .mu.g each of Cy3 and Cy5 labeled cDNA synthesis products in 5.times.SSC, 0.2% SDS hybridization buffer. The sample mixture is heated to 65.degree. C. for 5 minutes and is aliquoted onto the microarray surface and covered with an 1.8 cm.sup.2 coverslip. The arrays are transferred to a waterproof chamber having a cavity just slightly larger than a microscope slide. The chamber is kept at 100% humidity internally by the addition of 140 .mu.l of 5.times.SSC in a corner of the chamber. The chamber containing the arrays is incubated for about 6.5 hours at 60.degree. C. The arrays are washed for 10 min at 45.degree. C. in a first wash buffer (1.times.SSC, 0.1% SDS), three times for 10 minutes each at 45.degree. C. in a second wash buffer (0.1.times.SSC), and dried.

[0336] Detection

[0337] Reporter-labeled hybridization complexes are detected with a microscope equipped with an Innova 70 mixed gas 10 W laser (Coherent, Inc., Santa Clara Calif.) capable of generating spectral lines at 488 nm for excitation of Cy3 and at 632 nm for excitation of Cy5. The excitation laser light is focused on the array using a 20.times. microscope objective (Nikon, Inc., Melville N.Y.). The slide containing the array is placed on a computer-controlled X-Y stage on the microscope and raster-scanned past the objective. The 1.8 cm.times.1.8 cm array used in the present example is scanned with a resolution of 20 micrometers.

[0338] In two separate scans, a mixed gas multiline laser excites the two fluorophores sequentially. Emitted light is split, based on wavelength, into two photomultiplier tube detectors (PMT R1477, Hamamatsu Photonics Systems, Bridgewater N.J.) corresponding to the two fluorophores. Appropriate filters positioned between the array and the photomultiplier tubes are used to filter the signals. The emission maxima of the fluorophores used are 565 nm for Cy3 and 650 nm for Cy5. Each array is typically scanned twice, one scan per fluorophore using the appropriate filters at the laser source, although the apparatus is capable of recording the spectra from both fluorophores simultaneously.

[0339] The sensitivity of the scans is typically calibrated using the signal intensity generated by a cDNA control species added to the sample mixture at a known concentration. A specific location on the array contains a complementary DNA sequence, allowing the intensity of the signal at that location to be correlated with a weight ratio of hybridizing species of 1:100,000. When two samples from different sources (e.g., representing test and control cells), each labeled with a different fluorophore, are hybridized to a single array for the purpose of identifying genes that are differentially expressed, the calibration is done by labeling samples of the calibrating cDNA with the two fluorophores and adding identical amounts of each to the hybridization mixture.

[0340] The output of the photomultiplier tube is digitized using a 12-bit Rn-835H analog-to-digital (A/D) conversion board (Analog Devices, Inc., Norwood Mass.) installed in an IBM-compatible PC computer. The digitized data are displayed as an image where the signal intensity is mapped using a linear 20-color transformation to a pseudocolor scale ranging from blue (low signal) to red (high signal). The data is also analyzed quantitatively. Where two different fluorophores are excited and measured simultaneously, the data are first corrected for optical crosstalk (due to overlapping emission spectra) between the fluorophores using each fluorophore's emission spectrum.

[0341] A grid is superimposed over the fluorescence signal image such that the signal from each spot is centered in each element of the grid. The fluorescence signal within each element is then integrated to obtain a numerical value corresponding to the average intensity of the signal. The software used for signal analysis is the GEMTOOLS gene expression analysis program (Incyte).

[0342] XI. Complementary Polynucleotides

[0343] Sequences complementary to the PKIN-encoding sequences, or any parts thereof, are used to detect, decrease, or inhibit expression of naturally occurring PKIN. Although use of oligonucleotides comprising from about 15 to 30 base pairs is described, essentially the same procedure is used with smaller or with larger sequence fragments. Appropriate oligonucleotides are designed using OLIGO 4.06 software (National Biosciences) and the coding sequence of PKIN. To inhibit transcription, a complementary oligonucleotide is designed from the most unique 5' sequence and used to prevent promoter binding to the coding sequence. To inhibit translation, a complementary oligonucleotide is designed to prevent ribosomal binding to the PKIN-encoding transcript.

[0344] XII. Expression of PKIN

[0345] Expression and purification of PKIN is achieved using bacterial or virus-based expression systems. For expression of PKIN in bacteria, cDNA is subcloned into an appropriate vector containing an antibiotic resistance gene and an inducible promoter that directs high levels of cDNA transcription. Examples of such promoters include, but are not limited to, the trp-lac (tac) hybrid promoter and the T5 or T7 bacteriophage promoter in conjunction with the lac operator regulatory element. Recombinant vectors are transformed into suitable bacterial hosts, e.g., BL21(DE3). Antibiotic resistant bacteria express PKIN upon induction with isopropyl beta-D-thiogalactopyranoside (IPTG). Expression of PKIN in eukaryotic cells is achieved by infecting insect or mammalian cell lines with recombinant Autographica californica nuclear polyhedrosis virus (AcMNPV), commonly known as baculovirus. The nonessential polyhedrin gene of baculovirus is replaced with cDNA encoding PKIN by either homologous recombination or bacterial-mediated transposition involving transfer plasmid intermediates. Viral infectivity is maintained and the strong polyhedrin promoter drives high levels of cDNA transcription. Recombinant baculovirus is used to infect Spodoptera frugiperda (Sf9) insect cells in most cases, or human hepatocytes, in some cases. Infection of the latter requires additional genetic modifications to baculovirus. (See Engelhard, E. K. et al. (1994) Proc. Natl. Acad. Sci. USA 91:3224-3227; Sandig, V. et al. (1996) Hum. Gene Ther. 7:1937-1945.)

[0346] In most expression systems, PKIN is synthesized as a fusion protein with, e.g., glutathione S-transferase (GST) or a peptide epitope tag, such as FLAG or 6-His, permitting rapid, single-step, affinity-based purification of recombinant fusion protein from crude cell lysates. GST, a 26-kilodalton enzyme from Schistosoma janonicum, enables the purification of fusion proteins on immobilized glutathione under conditions that maintain protein activity and antigenicity (Amersham Pharmacia Biotech). Following purification, the GST moiety can be proteolytically cleaved from PKIN at specifically engineered sites. FLAG, an 8-amino acid peptide, enables immunoaffinity purification using commercially available monoclonal and polyclonal anti-FLAG antibodies (Eastman Kodak). 6-His, a stretch of six consecutive histidine residues, enables purification on metal-chelate resins (QIAGEN). Methods for protein expression and purification are discussed in Ausubel (1995, supra, ch. 10 and 16). Purified PKIN obtained by these methods can be used directly in the assays shown in Examples XVI, XVII, and XVIII where applicable.

[0347] XIII. Functional Assays

[0348] PKIN function is assessed by expressing the sequences encoding PKIN at physiologically elevated levels in mammalian cell culture systems. cDNA is subcloned into a mammalian expression vector containing a strong promoter that drives high levels of cDNA expression. Vectors of choice include PCMV SPORT (Life Technologies) and PCR3.1 (Invitrogen, Carlsbad Calif.), both of which contain the cytomegalovirus promoter. 5-10 .mu.g of recombinant vector are transiently transfected into a human cell line, for example, an endothelial or hematopoietic cell line, using either liposome formulations or electroporation. 1-2 .mu.g of an additional plasmid containing sequences encoding a marker protein are co-transfected. Expression of a marker protein provides a means to distinguish transfected cells from nontransfected cells and is a reliable predictor of cDNA expression from the recombinant vector. Marker proteins of choice include, e.g., Green Fluorescent Protein (GFP; Clontech), CD64, or a CD64-GFP fusion protein. Flow cytometry (FCM), an automated, laser optics-based technique, is used to identify transfected cells expressing GFP or CD64GFP and to evaluate the apoptotic state of the cells and other cellular properties. FCM detects and quantifies the uptake of fluorescent molecules that diagnose events preceding or coincident with cell death. These events include changes in nuclear DNA content as measured by staining of DNA with propidium iodide; changes in cell size and granularity as measured by forward light scatter and 90 degree side light scatter; down-regulation of DNA synthesis as measured by decrease in bromodeoxyuridine uptake; alterations in expression of cell surface and intracellular proteins as measured by reactivity with specific antibodies; and alterations in plasma membrane composition as measured by the binding of fluorescein-conjugated Annexin V protein to the cell surface. Methods in flow cytometry are discussed in Ormerod, M. G. (1994) Flow Cytometry, Oxford, New York N.Y.

[0349] The influence of PKIN on gene expression can be assessed using highly purified populations of cells transfected with sequences encoding PKIN and either CD64 or CD64-GFP. CD64 and CD64-GFP are expressed on the surface of transfected cells and bind to conserved regions of human immunoglobulin G (IgG). Transfected cells are efficiently separated from nontransfected cells using magnetic beads coated with either human IgG or antibody against CD64 (DYNAL, Lake Success N.Y.). mRNA can be purified from the cells using methods well known by those of skill in the art. Expression of mRNA encoding PKIN and other genes of interest can be analyzed by northern analysis or microarray techniques.

[0350] XIV. Production of PKJIN Specific Antibodies

[0351] PKJN substantially purified using polyacrylamide gel electrophoresis (PAGE; see, e.g., Harrington, M. G. (1990) Methods Enzymol. 182:488-495), or other purification techniques, is used to immunize rabbits and to produce antibodies using standard protocols.

[0352] Alternatively, the PKIN amino acid sequence is analyzed using LASERGENE software (DNASTAR) to determine regions of high immunogenicity, and a corresponding oligopeptide is synthesized and used to raise antibodies by means known to those of skill in the art. Methods for selection of appropriate epitopes, such as those near the C-terminus or in hydrophilic regions are well described in the art. (See, e.g., Ausubel, 1995, supra, ch. 11.)

[0353] Typically, oligopeptides of about 15 residues in length are synthesized using an ABI 431A peptide synthesizer (Applied Biosystems) using FMOC chemistry and coupled to KLH (Sigma-Aldrich, St. Louis Mo.) by reaction with N-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) to increase immunogenicity. (See, e.g., Ausubel, 1995, supra.) Rabbits are immunized with the oligopeptide-KLH complex in complete Freund's adjuvant. Resulting antisera are tested for antipeptide and anti-PKIN activity by, for example, binding the peptide or PKIN to a substrate, blocking with 1% BSA, reacting with rabbit antisera, washing, and reacting with radio-iodinated goat anti-rabbit IgG.

[0354] XV. Purification of Naturally Occurring PUIN Using Specific Antibodies

[0355] Naturally occurring or recombinant PKIN is substantially purified by immunoaffinity chromatography using antibodies specific for PKIN. An immunoaffinity column is constructed by covalently coupling anti-PKIN antibody to an activated chromatographic resin, such as CNBr-activated SEPHAROSE (Amersham Pharmacia Biotech). After the coupling, the resin is blocked and washed according to the manufacturer's instructions.

[0356] Media containing PKIN are passed over the immunoaffinity column, and the column is washed under conditions that allow the preferential absorbance of PKIN (e.g., high ionic strength buffers in the presence of detergent). The column is eluted under conditions that disrupt antibody/PKIN binding (e.g., a buffer of pH 2 to pH 3, or a high concentration of a chaotrope, such as urea or thiocyanate ion), and PKIN is collected.

[0357] XVI. Identification of Molecules which Interact with PKIN

[0358] PKIN, or biologically active fragments thereof, are labeled with .sup.125I Bolton-Hunter reagent. (See, e.g., Bolton A. E. and W. M. Hunter (1973) Biochem. J. 133:529-539.) Candidate molecules previously arrayed in the wells of a multi-well plate are incubated with the labeled PKIN, washed, and any wells with labeled PKIN complex are assayed. Data obtained using different concentrations of PKIN are used to calculate values for the number, affinity, and association of PKIN with the candidate molecules.

[0359] Alternatively, molecules interacting with PKIN are analyzed using the yeast two-hybrid system as described in Fields, S. and O. Song (1989) Nature 340:245-246, or using commercially available kits based on the two-hybrid system, such as the MATCHAE system (Clontech).

[0360] PKIN may also be used in the PATHCALLING process (CuraGen Corp., New Haven Conn.) which employs the yeast two-hybrid system in a high-throughput manner to determine all interactions between the proteins encoded by two large libraries of genes (Nandabalan, K. et al. (2000) U.S. Pat. No. 6,057,101).

[0361] XVIII. Demonstration of PKIN Activity

[0362] Generally, protein kinase activity is measured by quantifying the phosphorylation of a protein substrate by PKIN in the presence of gamma-labeled .sup.32P-ATP. PKIN is incubated with the protein substrate, .sup.32P-ATP, and an appropriate kinase buffer. The .sup.32P incorporated into the substrate is separated from free .sup.32P-ATP by electrophoresis and the incorporated .sup.32P is counted using a radioisotope counter. The amount of incorporated .sup.32P is proportional to the activity of PKIN. A determination of the specific amino acid residue phosphorylated is made by phosphoamino acid analysis of the hydrolyzed protein.

[0363] In one alternative, protein kinase activity is measured by quantifying the transfer of gamma phosphate from adenosine triphosphate (ATP) to a serine, threonine or tyrosine residue in a protein substrate. The reaction occurs between a protein kinase sample with a biotinylated peptide substrate and gamma .sup.32P-ATP. Following the reaction, free avidin in solution is added for binding to the biotinylated .sup.32P-peptide product. The binding sample then undergoes a centrifugal ultrafiltration process with a membrane which will retain the product-avidin complex and allow passage of free gamma .sup.32P-ATP. The reservoir of the centrifuged unit containing the .sup.32P-peptide product as retentate is then counted in a scintillation counter. This procedure allows assay of any type of protein kinase sample, depending on the peptide substrate and kinase reaction buffer selected. This assay is provided in kit form (ASUA, Affinity Ultrafiltration Separation Assay, Transbio Corporation, Baltimore Md., U.S. Pat. No. 5,869,275). Suggested substrates and their respective enzymes are as follows: Histone HI (Sigma) and p34.sup.cdc2kinase, Annexin I, Angiotensin (Sigma) and EGF receptor kinase, Annexin II and src kinase, ERK1 & ERK2 substrates and MEK, and myelin basic protein and ERK (Pearson, J. D. et al. (1991) Methods Enzymol. 200:62-81).

[0364] In another alternative, protein kinase activity of PKIN is demonstrated in vitro in an assay containing PKIN, 50 .mu.l of kinase buffer, 1 .mu.g substrate, such as myelin basic protein (MBP) or synthetic peptide substrates, 1 mM DTT, 10 .mu.g ATP, and 0.5 .mu.Ci [.gamma.-.sup.32P]ATP. The reaction is incubated at 30.degree. C. for 30 minutes and stopped by pipetting onto P81 paper. The unincorporated [.gamma.-.sup.32P]ATP is removed by washing and the incorporated radioactivity is measured using a radioactivity scintillation counter. Alternatively, the reaction is stopped by heating to 100.degree. C. in the presence of SDS loading buffer and visualized on a 12% SDS polyacrylamide gel by autoradiography. Incorporated radioactivity is corrected for reactions carried out in the absence of PKIN or in the presence of the inactive kinase, K38A.

[0365] In yet another alternative, adenylate kinase or guanylate kinase activity may be measured by the incorporation of .sup.32P from gamma-labeled .sup.32P-ATP into ADP or GDP using a gamma radioisotope counter. The enzyme, in a kinase buffer, is incubated together with the appropriate nucleotide mono-phosphate substrate (AMP or GMP) and .sup.32P-labeled ATP as the phosphate donor. The reaction is incubated at 37.degree. C. and terminated by addition of trichloroacetic acid. The acid extract is neutralized and subjected to gel electrophoresis to separate the mono-, di-, and triphosphonucleotide fractions. The diphosphonucleotide fraction is cut out and counted. The radioactivity recovered is proportional to the enzyme activity.

[0366] In yet another alternative, other assays for PKIN include scintillation proximity assays (SPA), scintillation plate technology and filter binding assays. Useful substrates include recombinant proteins tagged with glutathione transferase, or synthetic peptide substrates tagged with biotin. Inhibitors of PKIN activity, such as small organic molecules, proteins or peptides, may be identified by such assays.

[0367] XVII. Enhancement/Inhibition of Protein Kinase Activity

[0368] Agonists or antagonists of PKIN activation or inhibition may be tested using assays described in section XVII. Agonists cause an increase in PKIN activity and antagonists cause a decrease in PKIN activity.

[0369] Various modifications and variations of the described methods and systems of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with certain embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to those skilled in molecular biology or related fields are intended to be within the scope of the following claims.

2TABLE 1 Poly- peptide Incyte Incyte SEQ ID Incyte Polynucleotide Polynucleotide Project ID NO: Polypeptide ID SEQ ID NO: ID 2890544 1 2890544CD1 19 2890544CB1 7472693 2 7472693CD1 20 7472693CB1 3107952 3 3107952CD1 21 3107952CB1 5544420 4 5544420CD1 22 5544420CB1 7472832 5 7472832CD1 23 7472832CB1 1551456 6 1551456CD1 24 1551456CB1 2589355 7 2589355CD1 25 2589355CB1 4357117 8 4357117CD1 26 4357117CB1 5511992 9 5511992CD1 27 5511992CB1 7474560 10 7474560CD1 28 7474560CB1 7474602 11 7474602CD1 29 7474602CB1 7475509 12 7475509CD1 30 7475509CB1 7475491 13 7475491CD1 31 7475491CB1 2192119 14 2192119CD1 32 2192119CB1 7474496 15 7474496CD1 33 7474496CB1 1834248 16 1834248CD1 34 1834248CB1 71584520 17 71584520CD1 35 71584520CB1 7475538 18 7475538CD1 36 7475538CB1

[0370]

3TABLE 2 Polypeptide Incyte GenBank Probability SEQ ID NO: Polypeptide ID ID NO: score GenBank Homolog 1 2890544CD1 g5305331 0 protein kinase Myak-L [Mus musculus] 2 7472693CD1 g790790 2.4e-144 cam kinase I [Homo sapiens] 3 3107952CD1 g1403532 2.3e-226 KIS protein kinase [Rattus norvegicus] (Maucuer, A. et al. (1997) J. Biol. Chem. 272: 23151-23156) 4 5544420CD1 g205278 6.4e-293 male germ cell-associated kinase (mak) [Rattus norvegicus] (Matsushime, H. et al. (1990) Mol. Cell. Biol. 10: 2261-2268) 5 7472832CD1 g438373 0.0 protein kinase C mu [Homo sapiens] (Johannes, F. J. et al. (1994) J. Biol. Chem. 269: 6140-6148) 6 1551456CD1 g4099088 3.8e-26 [Arabidopsis thaliana] SNF1 family protein kinase 7 2589355CD1 g6760436 1.2e-144 [Gallus gallus] gin-induced kinase 8 4357117CD1 g6552404 2.4e-199 [Rattus norvegicus] DLG6 alpha 9 5511992CD1 g971420 3.9e-231 mixed lineage kinase 2 [Homo sapiens] Dorow, D. S. et al., (1995) Eur. J. Biochem. 234: 492-500 9 5511992CD1 g12005724 0 [5' incom] [Homo sapiens] mixed lineage kinase MLK1 10 7474560CD1 g4691541 7.3e-102 Adenylate kinase 5 [Homo sapiens]. Van Rompay, A. R. et al. (1999) Identification of a novel human adenylate kinase cDNA cloning, expression analysis, chromosome localization and characterization of the recombinant protein, Eur. J. Biochem. 261: 509-516. 11 7474602CD1 g439614 8.9e-145 CaM-like protein kinase [Rattus norvegicus]. Cho, F. S. et al. (1994) Characterization of a rat cDNA clone encoding calcium/calmodulin- dependent protein kinase I, Biochim. Biophys. Acta 1224: 156-160. 12 7475509CD1 g28577 2.4e-112 Nucleoside-triphosphate-adenylate kinase [Homo sapiens]. Xu, G. et al. (1992) Characterization of human adenylate kinase 3 (AK3) cDNA and mapping of the AK3 pseudogene to an intron of the NF1 gene, Genomics 13: 537-542. 13 7475491CD1 g2257588 1.4e-210 PCTAIRE3 [Rattus rattus]. Hirose, T. et al. (1997) PCTAIRE 2, a Cdc2-related serine/threonine kinase, is predominantly expressed in terminally differentiated neurons, Eur. J. Biochem. 249: 481-488. 14 2192119CD1 g3880563 2.0e-121 Predicted using Genefinder; similar to serine/threonine kinase; cDNA [Caenorhabditis elegans]. The C. elegans Sequencing Consortium (1998) Science 282: 2012-2018. 14 2192119CD1 g10442581 0 105-kDa kinase-like protein [Mus musculus] Liu, S. C. H., et al., (2000) Biochim. Biophys. Acta 1517: 148-152 15 7474496CD1 g6066585 0.0 GCN2 eIF2alpha kinase [Mus musculus]. 16 1834248CD1 g7595802 1.40E-252 ELKL motif kinase 2 short form [Mus musculus] 17 71584520CD1 g3927912 3.9e-157 calmodulin binding protein kinase [Fugu rubripes] Cottage, A. et al. (1999) FEBS Lett. 443: 370-374 18 7475538CD1 g4090958 9.2e-85 cell cycle related kinase [Homo sapiens] 18 7475538CD1 g9664926 0 CDK-related protein kinase PNQLARE [Mus musculus]

[0371]

4TABLE 3 SEQ Incyte Amino Potential Potential Analytical ID Polypeptide Acid Phosphorylation Glycosylation Signature Sequences, Methods and NO: ID Residues Sites Sites Domains and Motifs Databases 1 2890544CD1 1210 S20 T107 T163 N157 N111 N133 Protein_Kinase Atp: L196-K219 MOTIFS S211 T422 T666 N149 N262 Protein_Kinase St: L311-L323 MOTIFS S843 S853 T907 N471 N566 Eukaryotic protein kinase domain: HMMER_PFAM S127 T212 T508 N570 N1009 Y190-P411 I492-V518 S29 S37 T87 N1045 Tyrosine kinase catalytic domain BLIMPS_PRINTS S113 S169 S211 signature S396 T441 T474 PD00109B: K305-L323 T643 S856 S910 PROTEIN KINASE NUCLEAR HOMEODOMAIN BLAST_PRODOM T912 T938 S967 INTERACTING HOMEOBOX DNABINDING Y459 T1057 SERINE/THREONINE S1008 S1138 SERINE/THREONINEPROTEIN: S1187 PD150874: G1030-L1210 PROTEIN KINASE DOMAIN: BLAST_DOMO DM00004.vertline.P14680.vertline.371-694: V192-P509 2 7472693CD1 357 S64 S102 T117 N225 N311 PROTEIN_KINASE DOMAIN BLAST_DOMO S154 S169 S251 N332 DM00004.vertline.S57347.v- ertline.21-266: E24-C270 S326 S343 Y116 Protein_Kinase_Atp: L29-K52 MOTIFS Y133 S11 T45 Protein_Kinase_St: I140-Y152 MOTIFS T269 signal_cleavage: M1-A40 SPSCAN Eukaryotic protein kinase domain HMMER_PFAM pkinase: F23-I279 Protein kinases signatures and PROFILESCAN profile: D120-D176 Tyrosine kinase catalytic domain BLIMPS_PRINTS signature PR00109A: M98-V111 PR00109B: Y134-Y152 PR00109D: V202-E224 PROTEIN KINASE CALMODULINBINDING I BLAST_PRODOM CALCIUM/ CALMODULIN DEPENDENT TYPE CAM TRANSFERASE SERINE/THREONINE PROTEIN PHOSPHORYLATION PD012137: W278-L322 3 3107952CD1 419 S67 S117 S181 N253 transmembrane domain; HMMER S215 S221 T244 V223-L241 S290 T390 Eukaryotic protein kinase domain: HMMER_PFAM W23-F304 Tyrosine kinase catalytic domain BLIMPS_PRINTS PR00109B: F148-W166 PR00109D: V223-V245 PR00109E: P273-A295 SERINE/THREONINE PROTEIN KINASE BLAST_PRODOM PD153748: S305-V344 SPLICING FACTOR LIKE PROTEIN BLAST_PRODOM PD072361: T320-G412 RIBONUCLEOPROTEIN REPEAT BLAST_DOMO DM00012.vertline.A48249.vertline.299-407: P319-Y404 PROTEIN KINASE DOMAIN BLAST_DOMO DM00004.vertline.P08414.vert- line.44-285: R74-I293 DM00004.vertline.P32485.vertline.24-292: S48-P289 DM00004.vertline.P496571.vertline.101-409: C108-L241 4 5544420CD1 624 T274 S424 S436 N24 N252 N352 Eukaryotic protein kinase domain HMMER_PFAM S496 S41 S179 N384 N449 Y4-F284 S204 S315 S406 N455 N543 Tyrosine kinase catalytic domain BLIMPS_PRINTS S451 S466 S500 N569 PR00109B: F115-C133 S530 T6 S53 PR00109D: I181-T203 S161 T218 S388 PR00109E: A253-A275 S391 S420 S463 Protein kinases signatures & profile: PROFILESCAN T475 T581 S587 V101-Q153 T594 Y15 Y76 Protein kinases ATP-binding region: MOTIFS T571 L10-K33 Serine/Threonine protein kinases: MOTIFS F121-C133 ATP/GTP-binding site motif A (P-loop): MOTIFS G16-523 KINASE SERINE/THREONINE MALE GERM BLAST_PRODOM CELL TRANSFERASE ATP BINDING PD024663: Q285-R624 KINASE SERINE/THREONINE ATPBINDING II BLAST_PRODOM PHOSPHORYLATION CASEIN ALPHA CHAIN PD002608: V160-F284 KINASE TRANSFERASE ATP BINDING BLAST_PRODOM SERINE/THREONINE PROSPHORYLATION RECEPTOR TYROSINE PROTEIN PD000001: P155-F284 PROTEIN KINASE DOMAIN BLAST_DOMO DM00004.vertline.Q04859.vertline.6-274: T6-A275 DM00004.vertline.I48733.vertline.6-274: T6-A275 DM00004.vertline.P43294.vertline.14-281: M7-A275 DM00004.vertline.Q00526.vertline.6-286: M7-F284 5 7472832CD1 878 S197 S396 S797 N187 N431 Eukaryotic protein kinase domain: HMMER_PFAM S75 S11 T130 N432 N454 I551-L807 T218 S225 S333 N473 N729 Phorbol esters/diacylglycerol binding HMMER_PFAM S353 S362 T398 H139-C188 S408 T573 T589 H265-C314 T812 T853 S145 PH domain: HMMER_PFAM S206 T247 S387 T398-V478 T391 T392 T412 Protein kinases signatures & profile: PROFILESCAN T434 S604 S641 L650-S706 S706 Y87 5 Phorbol esters/diacylglycerol binding PROFILESCAN F151-S214 C278-A339 Tyrosine kinase catalytic domain: BLIMPS_PRINTS PR00109B: H664-L682 PR00109D: L732-D754 Diacylglycerol/phorbol-esterase BLIMPS_PRINTS PR00008B: C152-G161 PR00008C: Q291-C302 PR00008D: H303-L315 KINASE PHORBOLESTER C MU BLAST_PRODOM SERINE/THREONINE NPKCMU ATP BINDING PD039353: D323-V478 PD031784: V45-P138 PD027900: L807-L878 KINASE PHORBOLESTER BINDING BLAST_PRODOM TRANSFERASE SERINE/THREONINE ZINC ATPBINDING C DUPLICATION PD000215: H265-C314 Protein kinases signatures and profile BLAST_PRODOM (protein_kinase_tyr.prf): R826-A877 PROTEIN_KINASE_DOMAIN BLAST_DOMO DM00004.vertline.A53215.vert- line.585-829: P553-V798 DM00004.vertline.I48719.vertline.591-8- 35: P553-V798 DM04692.vertline.A37237.vertline.1-676: V556-S771 M04692.vertline.P05773.vertline.1-672: V556-P845 Phorbol esters/diacylglycerol MOTIFS binding domain: H139-C188 H265-C314 Protein kinases ATP-binding region MOTIFS signature: L557-K580 Serine/Threonine protein kinases MOTIFS active-site signature: I670-L682 6 1551456CD1 440 S11 S175 T74 Eukaryotic protein kinase domain: HMMER-PFAM S85 S230 T297 L164-E331 S361 S365 S376 Tyrosine kinase signature: BLIMPS-PRINTS S396 T426 S11 A192-L210, S260-S282 S19 S131 S155 PROTEIN KINASE DOMAIN BLAST-DOMO T214 S251 Y68 DM00004.vertline.P34244.vertline.82-359: Y378 Y167-L324 Protein kinase motif: MOTIFS I198-L210 7 2589355CD1 923 Y104 Y676 T258 N112 N317 PROTEIN KINASE DOMAIN BLAST-DOMO S355 T481 S584 DM00004.vertline.P27448.vertline.58-297: G25-I259 T38 S125 S289 Serine/Threonine protein kinases MOTIFS T356 S391 T447 active-site signature S448 T455 T481 Protein _Kinase_St: I135-L147 S500 S512 T875 Eukaryotic protein kinase domain HMMER-PFAM S890 T17 S254 pkinase: S24-M268 S336 T417 S472 Protein kinases signatures and profile PROFILESCAN S531 S542 S551 protein_kinase_tyr.prf: Y90-G167 S564 S576 S667 Tyrosine kinase catalytic site BLIMPS-PRINTS PR00109: T93-A106, Y129-L147, L195-P217 8 4357117CD1 442 T16 T267 T270 N406 SH3 domain signature PR00452: BLIMPS-PRINTS S295 S361 S142 C147-R159, A115-Q130, D132-I141 S229 S178 S180 PROTEIN DOMAIN SH3 KINASE GUANYLATE BLAST-PRODOM T311 S408 S437 TRANSFERASE ATPBINDING REPEAT GMP Y294 Y304 Y346 MEMBRANE PD001338: T267-Q360 GUANYLATE KINASE DM00755.vertline.A57653.vertline.370- BLAST-DOMO 570: P228-P431 PDZ signaling molecule domain PDZ: HMMER-PFAM I3-V83 Guanylate kinase Guanylate_kin: HMMER-PFAM T268-Y372 Guanylate kinase protein BL00856: BLIMPS-BLOCKS V264-V284, L292-R339 9 5511992CD1 1046 T72 T112 S118 N813 N862 Receptor tyrosine kinase BL00240: BLIMPS-BLOCKS S223 S276 T294 N964 E290-V337, V337-I389 S603 S707 S770 Receptor tyrosine kinase BL00239: BLIMPS-BLOCKS S781 S814 S880 E181-P228, L232-I254, W291-R340, T913 S531 T777 L345-I389 S89 T135 T363 Receptor tyrosine kinase BL00790: BLIMPS-BLOCKS T394 T395 T436 I154-C207, S298-W330, L356-M404 S599 S606 S636 Protein kinase signature and profile PROFILESCAN T644 S808 S821 protein_kinase_tyr.prf: L232-T294 S834 T884 T919 Tyrosine kinase catalytic site BLIMPS-PRINTS S923 S966 S972 PR00109: M210-S223, D248-I266, S983 T984 Y325 G301-I311, S320-I342, C364-F386 SH3 domain signature PR00452: BLIMPS-PRINTS P55-A65, D69-K84, D91-N100, R102-R114 SH3 domain: P55-R114 HMMER-PFAM Eukaryotic protein kinase domain HMMER-PFAM pkinase: L134-L393 KINASE DOMAIN SH3 MIXED LINEAGE BLAST-PRODOM SERINE/THREONINE WITH LEUCINE ZIPPER PROLINE PD024997: I396-A741 PROTEIN KINASE DOMAIN BLAST-DOMO DM00004.vertline.A53800.vert- line.119-368: L136-F386 Protein kinases ATP-binding region MOTIFS signature Protein_Kinase_Atp: I140-K161 Serine/Threonine protein kinases MOTIFS active-site signature Protein_Kinase_St: I254-I266 10 7474560CD1 357 S23, T65, Y103, KINASE ADENYLATE TRANSFERASE, ATP- BLAST-PRODOM S124, T140, BINDING ATP/AMP TRANSPHOSPHORYLASE S142, S181, ISOENZYME PROTEIN 3D-STRUCTURE T185, S200, MITOCHONDRION: PD000657: I175-R294. S211, S213, ADENYLATE KINASE: DM00290.vertline.P00570.vertline.1- BLAST-DOMO S245, T288, 131: E165-R294. T304, Y316, Adenylate kinase protein: BL00113A: BLIMPS-BLOCKS T340, T350 F174-L190; BL00113B: H198-A241; BL00113C: G247-V261; BL00113D: S298-T328. Adenylate kinase signature: PR00094A: BLIMPS-PRINTS F174-C187; PR00094B: G202-S216; PR00094C: F252-G268; PR00094D: P300- Y315; PR00094E: R317-Q331. Uridine kinase signature: PR00988A: BLIMPS-PRINTS C170-C187 Shikimate kinase family: PR01100A: BLIMPS-PRINTS I173-E188. Adenylate Kinase: I9-Q20, F252-Q263. MOTIFS Adenylate kinase: I11-L88, I175-Q331. HMMER-PFAM Adenylate kinase signature PROFILESCAN (adenylate_kinase.prf): V229-A284. 11 7474602CD1 355 S11, T45, S64, N225, N311, PROTEIN KINASE DOMAIN: BLAST-DOMO S102, Y116, N332 DM00004.vertline.S57347.vertline.21-266: E24-C270. T117, Y133, Protein kinases ATP-binding region MOTIFS S154, S169, signature: L29-K52. S251, T269, Serine/Threonine protein kinases MOTIFS S326, S343 active-site signature: I140-Y152. Eukaryotic protein kinase domain: HMMER-PFAM F23-I279. Protein kinases signatures and profile PROFILESCAN (protein_kinase_tyr.prf): D120-D176. Tyrosine kinase catalytic domain: BLIMPS-PRINTS PR00109A: M98-V111; PR00109B: Y134- Y152; PR00109D: V202-E224. PROTEIN KINASE; CALMODULINBINDING I BLAST-PRODOM CALCIUM/CALMODULINDEPENDENT TYPE CAM TRANSFERASE SERINE/THREONINE- PROTEIN PHOSPHORYLATION: PD012137: W278-L322. 12 7475509CD1 224 S16, S75, T118, ADENYLATE KINASE: DM00290.vertline.P27144.vertline.1- BLAST-DOMO S124, S132, 125: M1-R126. T201, Adenylate kinase signature: PR00094A: BLIMPS-PRINTS V9-C22; PR00094B: G37-G51; PR0094C: W85-D101; PR00094D: Q159-Y174; PR00094E: D176-V190. Shikimate kinase family: PR01100A: A8- BLIMPS-PRINTS Q23; PR01100E: D107-S124 (p < 0.0028). Adenylate Kinase Motif: W85-Q96. MOTIFS KINASE ADENYLATE TRANSFERASE, ATP- BLAST-PRODOM BINDING ATP/AMP TRANSPHOSPHORYLASE ISOENZYME, PROTEIN 3D-STRUCTURE MITOCHONDRION: PD000657: I10-V190. Adenylate kinase: I10-V190 HMMER-PFAM Adenylate kinase protein: BL00113A: BLIMPS-BLOCKS V9-125; BL00113B: H33-E76; BL00113C: R80-L94; BL00113D: S132-D162. Adenylate kinase signature PROFILESCAN (adenylate_kinase.prf): V64-F116. 13 7475491CD1 502 S12, T20, S24, PROTEIN KINASE DOMAIN: BLAST-DOMO S60, S64, S87, DM00004.vertline.Q04899.vertline.122-392: V173-A444, T104, S122, KINASE SERINE/THREONINE PROTEIN BLAST-PRODOM S126, S156, TRANSFERASE ATP-BINDING DOMAIN- S160, Y183, PCTAIRE1, PCTAIRE2, PCTAIRE3, CRK5 T232, T239, ALTERNATIVE PD007333: D120-T171 T348, T364, Tyrosine kinase catalytic domain: BLIMPS-PRINTS T373, T387, PR00109B: Y283-I301 S455, S471, Protein kinases signatures and profile PROFILESCAN S500, (protein_kinase_tyr.prf): D235-A317 Eukaryotic protein kinase domain HMMER-PFAM (pkinase): Y172-F453, Protein kinases ATP-binding region MOTIFS signature: L178-K201 Serine/Threonine protein kinases MOTIFS active-site signature: I289-I301 14 2192119CD1 791 T39, T70, N447 PROTEIN KINASE DOMAIN BLAST-DOMO T85, S193, DM00004.vertline.P52304.vertline.27-267: S269, T319, K69-P254 (p > 1.3e-06) S342, S359, T361, T408, S462, S499, S521, S535, S539, T545, T581, T610, T617, S622, S662, S668, S677, S693, S716, S726, S737, Y184 15 7474496CD1 1651 T65, T82, N100, N245, PROTEIN KINASE DOMAIN: BLAST-DOMO S108, S144, 1057, 1197, DM0004.vertline.P15442.vertline.645-902: S732-A994, S210, T215, 1203, 1248, EUKARYOTIC INITIATION FACTOR KINASE BLAST-PRODOM S216, S247, 1416, 1520, (EIF2 ALPHA): PD156018: D13-D219 S250, T329, 1602 Tyrosine kinase catalytic domain: BLIMPS-PRINTS T346, T409, PR00109B: Y840-L858 S414, T441, Protein kinases signatures and profile PROFILESCAN S450, T478, (protein_kinase_tyr.prf): R826-A877 S553, S569, Eukaryotic protein kinase domain HMMER-PFAM T581, S690, (pkinase): S709, T725, K335-D446; F592-P664; Y799-L1003 S722, S732, Protein kinases ATP-binding region MOTIFS S733, S755, signature: L598-K621 S757, T811, Serine/Threonine protein kinases MOTIFS T824, S882, active-site signature: M846-L858 S917, T945, Eukaryotic protein kinase domain: HMMER-PFAM S947, S961, K335-D446; P504-I541; F592-P664; T993, T1022, Y799-L1003 S1038, S1059, Protein kinases signatures and profile PROFILE-SCAN T1062, T1182, (protein_kinase_tyr.prf): R826-A877 BLIMPS-PRINTS T1234, T1238, Tyrosine kinase catalytic domain: S1397, T1418, PR00109B: Y840-L858 S1437, S1457, Protein kinases ATP-binding region MOTIFS S1572, T1475, signature: L598-K621 S1547, T1548, Serine/Threonine protein kinases MOTIFS T1604, S1641, active-site signature: Y256, Y821, M846-L858 Y840, Y1626 16 1834248CD1 752 S139 S2 S210 N395 N532 Eukaryotic protein kinase domain: HMMER_PFAM S23 S27 S34 Y59-I310, S354 S399 S423 Tyrosine kinase catalytic domain BLIMPS_PRINTS S441 S458 S48 PR00109A: M135-V148 S494 S661 S666 PR00109B: Y171-L189 S710 T127 T281 PR00109D: V231-H259 T300 T323 T332 Protein kinases signatures and profile PROFILESCAN T344 T507 T511 protein_kinase_tyr.prf: Y132-S210 T515 T536 T564 Protein kinases ATP-binding region: MOTIFS T620 T624 T81 Protein_Kinase_Atp: I65-K88 Serine/Threonine protein kinases MOTIFS active-site Protein_Kinase_St: I177-L189 KINASE SERINE/THREONINE TRANSFERASE BLAST_PRODOM ATP BINDING PROTEIN EMK P78 CDC25C PD008571: S412-E632 KINASE SERINE/THREONINE TRANSFERASE BLAST_PRODOM ATP BINDING PROTEIN PAR1 KP78 EMK PD005838: I310-R410 KINASE SERINE/THREONINE TRANSFERASE BLAST_PRODOM ATP BINDING KIN1 EMK PAR1 PD004300: E650-L752 KINASE TRANSFERASE ATP BINDING BLAST_PRODOM SERINE/THREONINE PHOSPHORYLATION RECEPTOR TYROSINE PD000001: Y59-Y137 PROTEIN KINASE DOMAIN BLAST_DOMO DM00004.vertline.P27448.vertline.58-297: L61-L301 DM00004.vertline.I48609.vertline.55-294: L61-L301 DM00004.vertline.Q05512.vertline.55-294: L61-L301 DM00004.vertline.JC1446.vertline.20-261: R60-L301 17 71584520CD1 501 S118 S138 S292 Eukaryotic protein kinase domain HMMER_PFAM S341 S364 S482 pkinase: E37-I286 S483 S495 T103 Tyrosine kinase catalytic domain BLIMPS_PRINTS T21 T276 T422 PR00109B: Y135-Y153 T46 T470 T51 T7 PR00109D: V201-E223 T91 Y135 Y491 CALMODULIN BINDING PROTEIN BLAST_PRODOM PD059862:

G368-V443 PROTEIN KINASE CALMODULIN BINDING I BLAST_PRODOM CALCIUM/CALMODULIN DEPENDENT TYPE CAM TRANSFERASE SERINE/THREONINE PROTEIN PHOSPHORYLATION PD012137: W285-A335 CALMODULIN BINDING PROTEIN BLAST_PRODOM PD050813: M1-E34 PROTEIN KINASE DOMAIN BLAST_DOMO DM00004.vertline.S57347.vertline.21-266: D25-T276 DM00004.vertline.P08414.vertline.44-285: I38-T276 DM00004.vertline.P11798.vertline.15-261: C36-A277 DM00004.vertline.A44412.vertline.16-262: F35-A277 18 7475538CD1 346 S241 Y176 Y215 Eukaryotic protein kinase domain HMMER_PFAM pkinase: Y4-F288, Tyrosine kinase catalytic domain BLIMPS_PRINTS PR00109B: F117-I135 Protein kinases signatures and profile PROFILESCAN protein_kinase_tyr.prf: A69-D155 Protein kinases ATP-binding region MOTIFS signature Protein_Kinase_Atp: I10-K33 Serine/Threonine protein kinases MOTIFS active-site Protein_Kinase_St: I123-I135 KINASE TRANSFERASE SERINE/THREONINE BLAST_PRODOM ATP BINDING II PHOSPHORYLATION CASEIN ALPHA CHAIN PD002608: V164-F288 KINASE PROTEIN TRANSFERASE ATP BLAST_PRODOM BINDING SERINE/THREONINE PHOSPHORYLATION RECEPTOR TYROSINE TRANSMEMBRANE PD000001: Y169-P301 PROTEIN KINASE DOMAIN BLAST_DOMO DM00004.vertline.P29620.vertline.21-289: I10-A279 DM00004.vertline.Q00526.vertline.6-286: R9-F288 DM00004.vertline.P43450.vertline.6-276: R9-A279 DM00004.vertline.P23437.vertline.6-286: R9-F288

[0372]

5TABLE 4 Poly- Incyte nucleotide Polynucleotide Sequence Selected 5' 3' SEQ ID NO: ID Length Fragment(s) Sequence Fragments Position Position 19 2890544CB1 4224 1-1378, 4153-4224 6474032H1 (PLACFEB01) 545 1204 71089659V1 2981 3638 71083920V1 2936 3576 71083254V1 2263 2853 71084605V1 2351 2959 7286979H1 (BRAIFER06) 1 408 71254276V1 1091 1695 5980233F7 (MCLDTXT02) 3498 4224 71252928V1 1632 2197 7086636H1 (BRAUTDR03) 222 702 71082857V1 1680 2356 7313223H1 (LIVRFEE02) 498 928 20 7472693CB1 1736 72-149, 1337-1736 70520498D1 760 1354 609792R6 (COLNNOT01) 145 736 g1544947 1 385 70518493D1 716 1182 70518085D1 1172 1736 21 3107952CB1 1824 1802-1824 7308849H1 (MMLR1DT01) 1215 1807 2925973F6 (TLYMNOT04) 715 1215 3459433F6 (293TF1T01) 1 582 2927552F6 (TLYMNOT04) 1309 1824 3107952F6 (BRSTTUT15) 507 1161 22 5544420CB1 2201 1285-1629, 2128- 6922389H1 (PLACFER06) 1458 1597 2201, 256-376 7739285H1 (THYMNOE01) 646 834 5544420F6 (TESTNOC01) 807 1187 4206166F6 (BRONNOT02) 2041 2201 GNN: g5924006_004.edit 368 1871 GBI.g5924006.raw.comp 1 492 2512558F6 (LIVRTUT04) 1617 1720 g2882961 1644 2127 6909108J1 (PITUDIR01) 1138 1317 23 7472832CB1 2974 2933-2974, 1-654 6910588J1 (PITUDIR01) 1994 2577 2940460H1 (THYNFET02) 2682 2956 7274331H2 (KIDETXJ01) 1876 2503 60205600U1 559 1091 23 60205598U1 690 1339 6426807H1 (LUNGNON07) 2411 2951 3344032H1 (SPLNNOT09) 2708 2958 6811278J1 (SKIRNOR01) 1 647 6910588H1 (PITUDIR01) 1369 2003 g1860144 2512 2974 1732303F6 (BRSTTUT08) 1230 1731 24 1551456CB1 3648 1906-2052, 1-752 6830947J1 (SINTNOR01) 2357 3121 6834559H1 (BRSTNON02) 1610 2330 6808991J1 (SKIRNOR01) 222 921 1389125H1 (EOSINOT01) 1 246 1680356F6 (STOMFET01) 3139 3648 6870264H1 (BRAGNON02) 3374 3648 7714085J1 (SINTFEE02) 298 950 7746056H1 (ADRETUE04) 875 1491 3296183F6 (TLYJINT01) 1553 2310 70349297D1 3030 3648 70872871V1 953 1599 70875814V1 2218 2955 25 2589355CB1 4719 1835-1863, 6500833H1 (PROSTUS25) 2498 3226 1-1135, 2380-3186 2313925T6 (NGANNOT01) 3264 3885 6483636F9 (MIXDUNB01) 2716 3270 70985467V1 388 980 7251743H1 (PROSTMY01) 810 1299 6864125H1 (BPAGNON02) 1323 1998 6056925H1 (BRAENOT04) 1322 1735 6705826H1 (HEAADIR01) 4080 4719 7652463H2 (STOMTDE01) 997 1704 7039646H1 (UTRSTMR02) 1 566 7257374H1 (SKIRTDC01) 1977 2560 70402344D1 3105 3674 2586850F6 (BRAITUT22) 3840 4348 924331H1 (RATRNOT02) 4009 4386 26 4357117CB1 1651 1-267 GS.4357117.fasta 1 1329 6131509F6 (BMARTXT02) 1134 1651 27 5511992CB1 3141 1672-1833, GS.5511992.fasta 1 3141 71-157, 665-828, 1298-1538, 2580- 2878 28 7474560CB1 1244 1205-1244, 1-131 6311370H1 (NERDTDN03) 616 1236 6853555H1 (BRAIFEN08) 1 683 6997205H1 (BRAXTDR17) 691 1244 29 7474602CB1 1661 72-149 70518523D1 577 1171 6772112J1 (BRAUNOR01) 1 640 346275T7 (THYMNOT02) 1043 1661 6124350H1 (BRAHNON05) 634 1180 30 7475509CB1 912 881-912 944796H1 (RATRNOT02) 1 93 3616204F6 (EPIPNOT01) 69 699 71147872V1 193 912 31 7475491CB1 2858 1-269, 2768-2858 7232424H1 (BRAXTDR15) 1831 2429 7004062H1 (COLNFEC01) 1967 2495 7677070J1 (NOSETUE01) 1 564 7716833J1 (SINTFEE02) 1293 1877 754239R6 (BRAITUT02) 1000 1575 7067423H1 (BRATNOR01) 499 1106 7428450H1 (UTRMTMR02) 2179 2858 70680595V1 654 1111 32 2192119CB1 2817 1-249 1687835F6 (PROSTUT10) 2416 2817 6769461J1 (BRAUNOR01) 221 385 1819105F6 (PROSNOT20) 1052 1721 7716523H1 (SINTFEE02) 498 1072 1796441T6 (PROSTUT05) 2186 2793 1618475T6 (BRAITUT12) 2217 2783 7675143H1 (NOSETUE01) 1130 1739 6145288H1 (BRANDIT03) 1543 2170 3189653H1 (THYMNON04) 1855 2196 7646563H1 (UTRSTUE01) 367 1049 g6700560 1 393 33 7474496CB1 5305 3463-3753, 1403- 70886570V1 3332 3875 1781, 2384-2464, 1-644, 5126-5305 33 429360R6 (BLADNOT01) 2735 3311 70885734V1 4100 4744 1830377F6 (THP1AZT01) 3649 4093 6488464H1 (MIXDUNB01) 2852 3473 488190R6 (HNT2AGT01) 2317 2865 70888476V1 4651 5305 1832566R6 (BRAINON01) 4034 4547 34 1834248CB1 3269 1754-1773, 1-142, GNN.g7139831_000025_004 1293 2275 3072-3269 6146293H1 (BRANDIT03) 1791 2354 6272238H2 (BRAIFEN03) 2635 3263 6893004J1 (BRAITDR03) 684 1248 7663341J1 (UTRSTME01) 873 1492 4001427R6 (HNT2AZS07) 2927 3264 60202068B1 2368 2916 60202069B1 2334 2864 6954283H1 (BRAITDR02) 1 718 g810284 2771 3269 35 71584520CB1 3017 2968-3017, 1-49, 1287320T6 (BRAINOT11) 2615 2983 1234-1322, 679-736 71579751V1 1083 1791 71580777V1 586 1149 6764749H1 (BRAUNOR01) 1406 2001 7581090H1 (BRAIFEC01) 1 577 1295833H1 (PGANNOT03) 2783 3017 1414795F6 (BRAINOT12) 2183 2508 7362189H1 (BRAIFEE05) 458 1003 2157112T6 (BRAINOT09) 2388 2977 2157112F6 (BRAINOT09) 1963 2450 36 7475538CB1 2168 811-870, 933-1227 6855691H1 (BRAIFEN08) 1500 2168 70644867V1 613 1261 70645804V1 515 1249 70645323V1 1 595 71564044V1 1277 1993 71565564V1 1156 1924

[0373]

6TABLE 5 Poly- Incyte nucleotide Project SEQ ID NO: ID Representative Library 19 2890544CB1 MCLDTXT02 20 7472693CB1 COLNNOT01 21 3107952CB1 TLYMNOT04 22 5544420CB1 SEMVNOT01 23 7472832CB1 SKIRNOR01 24 1551456CB1 LVENNOT03 25 2589355CB1 BRADDIR01 26 4357117CB1 BMARTXT02 27 5511992CB1 SINTFEE02 28 7474560CB1 BRAYDIN03 29 7474602CB1 COLNNOT01 30 7475509CB1 BRAITVT21 31 7475491CB1 SCOMDIT01 32 2192119CB1 PROSTUS23 33 7474496CB1 BRAINON01 34 1834248CB1 BRAITUT22 35 71584520CB1 BRAIFEC01 36 7475538CB1 BRAIFEN08

[0374]

7TABLE 6 Library Vector Library Description BMARTXT02 pINCY Library was constructed using RNA isolated from treated SH-SY5Y cell line derived from bone marrow neuroblastoma tumor cells removed from a 4-year-old Caucasian female. The cells were cultured in the presence of retinoic acid. BRADDIR01 pINCY Library was constructed using RNA isolated from diseased choroid plexus tissue of the lateral ventricle, removed from the brain of a 57-year-old Caucasian male, who died from a cerebrovascular accident. BRAIFEC01 pINCY This large size-fractionated library was constructed using RNA isolated from brain tissue removed from a Caucasian male fetus who was stillborn with a hypoplastic left heart at 23 weeks' gestation. BRAIFEN08 pINCY This normalized fetal brain tissue library was constructed from 400 thousand independent clones from a fetal brain tissue library. Starting RNA was made from brain tissue removed from a Caucasian male fetus who was stillborn with a hypoplastic left heart at 23 weeks' gestation. The library was normalized in 2 rounds using conditions adapted from Soares et al., PNAS (1994) 91: 9228 and Bonaldo et al., Genome Research (1996) 6: 791, except that a significantly longer (48 hours/round) reannealing hybridization was used. BRAINON01 PSPORT1 Library was constructed and normalized from 4.88 million independent clones from a brain tissue library. RNA was made from brain tissue removed from a 26-year-old Caucasian male during cranioplasty and excision of a cerebral meningeal lesion. Pathology for the associated tumor tissue indicated a grade 4 oligoastrocytoma in the right fronto-parietal part of the brain. BRAITUT21 pINCY Library was constructed using RNA isolated from brain tumor tissue removed from the midline frontal lobe of a 61-year-old Caucasian female during excision of a cerebral meningeal lesion. Pathology indicated subfrontal meningothelial meningioma with no atypia. One ethmoid and mucosal tissue sample indicated meningioma. Family history included cerebrovascular disease, senile dementia, hyperlipidemia, benign hypertension, atherosclerotic coronary artery disease, congestive heart failure, and breast cancer. BRAITUT22 pINCY Library was constructed using RNA isolated from brain tumor tissue removed from the right frontal/parietal lobe of a 76-year-old Caucasian female during excision of a cerebral meningeal lesion. Pathology indicated a meningioma. Family history included senile dementia. BRAYDIN03 pINCY This normalized brain tissue library was constructed from 6.7 million independent clones from the BRAYDIT01 tissue library. Starting RNA was made from RNA isolated from diseased hypothalamus tissue removed from a 57-year- old Caucasian male who died from a cerebrovascular accident. Patient history included Huntington's disease and emphysema. The library was normalized in 2 rounds using conditions adapted from Soares et al., PNAS (1994) 91: 9228 and Bonaldo et al., Genome Research (1996) 6: 791, except that a significantly longer (48-hours/round) reannealing hybridization was used. The library was linearized and recircularized to select for insert containing clones. COLNNOT01 PSPORT1 Library was constructed using RNA isolated from colon tissue removed from a 75-year-old Caucasian male during a hemicolectomy. LVENNOT03 PSPORT1 Library was constructed using RNA isolated from the left ventricle tissue of a 31-year-old male. MCLDTXT02 pINCY Library was constructed using RNA isolated from treated umbilical cord blood dendritic cells removed from a male. The cells were treated with granulocyte/macrophage colony stimulating factor (GM-CSF), tumor necrosis factor alpha (TNF alpha), stem cell factor (SCF), phorbol myristate acetate (PMA), and ionomycin. The GM-CSF was added at time 0 at 100 ng/ml, the TNF alpha was added at time 0 at 2.5 ng/ml, the SCF was added at time 0 at 25 ng/ml. The RNA and ionomycin were added at 13 days for five hours. Incubation time was 13 days. PROSTUS23 pINCY This subtracted prostate tumor library was constructed using 10 million clones from a pooled prostate tumor library that was subjected to 2 rounds of subtractive hybridization with 10 million clones from a pooled prostate tissue library. The starting library for subtraction was constructed by pooling equal numbers of clones from 4 prostate tumor libraries using mRNA isolated from prostate tumor removed from Caucasian males at ages 58 (A), 61 (B), 66 (C), and 68 (D) during prostatectomy with lymph node excision. Pathology indicated adenocarcinoma in all donors. History included elevated PSA, induration and tobacco abuse in donor A; elevated PSA, induration, prostate hyperplasia, renal failure, osteoarthritis, renal artery stenosis, benign HTN, thrombocytopenia, hyperlipidemia, tobacco/alcohol abuse and hepatitis C (carrier) in donor B; elevated PSA, induration, and tobacco abuse in donor C; and elevated PSA, induration, hypercholesterolemia, and kidney calculus in donor D. The hybridization probe for subtraction was constructed by pooling equal numbers of cDNA clones from 3 prostate tissue libraries derived from prostate tissue, prostate epithelial cells, and fibroblasts from prostate stroma from 3 different donors. Subtractive hybridization conditions were based on the methodologies of Swaroop et al., NAR 19 (1991): 1954 and Bonaldo, et al. Genome Research 6 (1996): 791. SCOMDIT01 pINCY Library was constructed using RNA isolated from diseased spinal cord tissue removed from the base of the medulla of a 57-year-old Caucasian male who died from a cerebrovascular accident. Patient history included Huntington's disease and emphysema. SEMVNOT01 pINCY Library was constructed using RNA isolated from seminal vesicle tissue removed from a 58-year-old Caucasian male during radical prostatectomy. Pathology for the associated tumor tissue indicated adenocarcinoma (Gleason grade 3 + 2) of the prostate. Adenofibromatous hyperplasia was also present. The patient presented with elevated prostate specific antigen (PSA). Family history included a malignant breast neoplasm. SINTFEE02 PCDNA2.1 This 5' biased random primed library was constructed using RNA isolated from small intestine tissue removed from a Caucasian male fetus who died from Patau's syndrome (trisomy 13) at 20-weeks' gestation. Serology was negative. SKIRNOR01 PCDNA2.1 This random primed library was constructed using RNA isolated from skin tissue removed from the breast of a 17-year-old Caucasian female during bilateral reduction mammoplasty. Patient history included breast hypertrophy. Family history included benign hypertension. TLYMNOT04 pINCY Library was constructed using RNA isolated from activated Th1 cells. These cells were differentiated from umbilical cord CD4 T cells with IL-12 and B7- transfected COS cells, and then activated for six hours with anti-CD3 and anti-CD28 antibodies.

[0375]

8TABLE 7 Program Description Reference Parameter Threshold ABI A program that removes vector sequences and Applied Biosystems, Foster City, CA. FACTURA masks ambiguous bases in nucleic acid sequences. ABI/ A Fast Data Finder useful in comparing and Applied Biosystems, Foster City, CA; Mismatch <50% PARACEL annotating amino acid or nucleic acid sequences. Paracel Inc., Pasadena, CA. FDF ABI A program that assembles nucleic acid sequences. Applied Biosystems, Foster City, CA. AutoAssembler BLAST A Basic Local Alignment Search Tool useful in Altschul, S. F. et al. (1990) J. Mol. Biol. ESTs: Probability value = sequence similarity search for amino acid and 215: 403-410; Altschul, S. F. et al. (1997) 1.0E-8 or less nucleic acid sequences. BLAST includes five Nucleic Acids Res. 25: 3389-3402. Full Length sequences: functions: blastp, blastn, blastx, tblastn, and tblastx. Probability value = 1.0E-10 or less FASTA A Pearson and Lipman algorithm that searches for Pearson, W. R. and D. J. Lipman (1988) ESTs: fasta E value = 1.06E-6 similarity between a query sequence and a group of Proc. Natl. Acad Sci. USA 85: 2444-2448; Assembled ESTs: fasta sequences of the same type. FASTA comprises as Pearson, W. R. (1990) Methods Enzymol. Identity = 95% or greater and least five functions: fasta, tfasta, fastx, tfastx, and 183: 63-98; and Smith, T. F. and M. S. Match length = 200 bases or ssearch. Waterman (1981) Adv. Appl. Math. 2: greater; fastx E value = 1.0E-8 482-489. or less Full Length sequences: fastx score = 100 or greater BLIMPS A BLocks IMProved Searcher that matches a Henikoff, S. and J. G. Henikoff (1991) Probability value = 1.0E-3 or sequence against those in BLOCKS, PRINTS, Nucleic Acids Res. 19: 6565-6572; less DOMO, PRODOM, and PFAM databases to search Henikoff, J. G. and S. Henikoff (1996) for gene families, sequence homology, and structural Methods Enzymol. 266: 88-105; and fingerprint regions. Attwood, T. K. et al. (1997) J. Chem. Inf. Comput. Sci. 37: 417-424. HMMER An algorithm for searching a query sequence against Krogh, A. et al. (1994) J. Mol. Biol. PFAM hits: Probability value = hidden Markov model (HMM)-based databases of 235: 1501-1531; Sonnhammer, E. L. L. et 1.0E-3 or less protein family consensus sequences, such as PFAM. al. (1988) Nucleic Acids Res. 26: 320-322; Signal peptide hits: Score = 0 or Durbin, R. et al. (1998) Our World View, in greater a Nutshell, Cambridge Univ. Press, pp. 1-350. ProfileScan An algorithm that searches for structural and sequence Gribskov, M. et al. (1988) CABIOS 4: Normalized quality score .gtoreq. motifs in protein sequences that match sequence 61-66; Gribskov, M. et al. (1989) Methods GCG-specified "HIGH" value patterns defined in Prosite. Enzymol. 183: 146-159; Bairoch, A. et al. for that particular Prosite motif. (1997) Nucleic Acids Res. 25: 217-221. Generally, score = 1.4-2.1. Phred A base-calling algorithm that examines automated Ewing, B. et al. (1998) Genome Res. sequencer traces with high sensitivity and probability. 8: 175-185; Ewing, B. and P. Green (1998) Genome Res. 8: 186-194. Phrap A Phils Revised Assembly Program including SWAT Smith, T. F. and M. S. Waterman (1981) Score = 120 or greater; and CrossMatch, programs based on efficient Adv. Appl. Math. 2: 482-489; Smith, T. F. Match length = 56 or greater implementation of the Smith-Waterman algorithm, and M. S. Waterman (1981) J. Mol. Biol. useful in searching sequence homology and assembling 147: 195-197; and Green, P., University of DNA sequences. Washington, Seattle, WA. Consed A graphical tool for viewing and editing Phrap Gordon, D. et al. (1998) Genome Res. 8: assemblies. 195-202. SPScan A weight matrix analysis program that scans protein Nielson, H. et al. (1997) Protein Engineering Score = 3.5 or greater sequences for the presence of secretory signal peptides. 10: 1-6; Claverie, J. M. and S. Audic (1997) CABIOS 12: 431-439. TMAP A program that uses weight matrices to delineate Persson, B. and P. Argos (1994) J. Mol. transmembrane segments on protein sequences and Biol. 237: 182-192; Persson, B. and P. determine orientation. Argos (1996) Protein Sci. 5: 363-371. TMHMMER A program that uses a hidden Markov model (HMM) Sonnhammer, E. L. et al. (1998) Proc. Sixth to delineate transmembrane segments on protein Intl. Conf. on Intelligent Systems for Mol. sequences and determine orientation. Biol., Glasgow et al., eds., The Am. Assoc. for Artificial Intelligence Press, Menlo Park, CA, pp. 175-182. Motifs A program that searches amino acid sequences for Bairoch, A. et al. (1997) Nucleic Acids Res. patterns that matched those defined in Prosite. 25: 217-221; Wisconsin Package Program Manual, version 9, page M51-59, Genetics Computer Group, Madison, WI.

[0376]

Sequence CWU 1

1

36 1 1210 PRT Homo sapiens misc_feature Incyte ID No 2890544CD1 1 Met Ala Ser Gln Leu Gln Val Phe Ser Pro Pro Ser Val Ser Ser 1 5 10 15 Ser Ala Phe Cys Ser Ala Lys Lys Leu Lys Ile Glu Pro Ser Gly 20 25 30 Trp Asp Val Ser Gly Gln Ser Ser Asn Asp Lys Tyr Tyr Thr His 35 40 45 Ser Lys Thr Leu Pro Ala Thr Gln Gly Gln Ala Asn Ser Ser His 50 55 60 Gln Val Ala Asn Phe Asn Ile Pro Ala Tyr Asp Gln Gly Leu Leu 65 70 75 Leu Pro Ala Pro Ala Val Glu His Ile Val Val Thr Ala Ala Asp 80 85 90 Ser Ser Gly Ser Ala Ala Thr Ser Thr Phe Gln Ser Ser Gln Thr 95 100 105 Leu Thr His Arg Ser Asn Val Ser Leu Leu Glu Pro Tyr Gln Lys 110 115 120 Cys Gly Leu Lys Arg Lys Ser Glu Glu Val Asp Ser Asn Gly Ser 125 130 135 Val Gln Ile Ile Glu Glu His Pro Pro Leu Met Leu Gln Asn Arg 140 145 150 Thr Val Val Gly Ala Ala Ala Thr Thr Thr Thr Val Thr Thr Lys 155 160 165 Ser Ser Ser Ser Ser Gly Glu Gly Asp Tyr Gln Leu Val Gln His 170 175 180 Glu Ile Leu Cys Ser Met Thr Asn Ser Tyr Glu Val Leu Glu Phe 185 190 195 Leu Gly Arg Gly Thr Phe Gly Gln Val Ala Lys Cys Trp Lys Arg 200 205 210 Ser Thr Lys Glu Ile Val Ala Ile Lys Ile Leu Lys Asn His Pro 215 220 225 Ser Tyr Ala Arg Gln Gly Gln Ile Glu Val Ser Ile Leu Ser Arg 230 235 240 Leu Ser Ser Glu Asn Ala Asp Glu Tyr Asn Leu Val Arg Ser Tyr 245 250 255 Glu Cys Phe Gln His Lys Asn His Thr Cys Leu Val Phe Glu Met 260 265 270 Leu Glu Gln Asn Leu Tyr Asp Phe Leu Lys Gln Asn Lys Phe Ser 275 280 285 Pro Leu Pro Leu Lys Tyr Ile Arg Pro Ile Leu Gln Gln Val Ala 290 295 300 Thr Ala Leu Met Lys Leu Lys Ser Leu Gly Leu Ile His Ala Asp 305 310 315 Leu Lys Pro Glu Asn Ile Met Leu Val Asp Pro Val Arg Gln Pro 320 325 330 Tyr Arg Val Lys Val Ile Asp Phe Gly Ser Ala Ser His Val Ser 335 340 345 Lys Ala Val Cys Ser Thr Tyr Leu Gln Ser Arg Tyr Tyr Arg Ala 350 355 360 Pro Glu Ile Ile Leu Gly Leu Pro Phe Cys Glu Ala Ile Asp Met 365 370 375 Trp Ser Leu Gly Cys Val Ile Ala Glu Leu Phe Leu Gly Trp Pro 380 385 390 Leu Tyr Pro Gly Ala Ser Glu Tyr Asp Gln Ile Arg Tyr Ile Ser 395 400 405 Gln Thr Gln Gly Leu Pro Ala Glu Tyr Leu Leu Ser Ala Gly Thr 410 415 420 Lys Thr Thr Arg Phe Phe Asn Arg Asp Pro Asn Leu Gly Tyr Pro 425 430 435 Leu Trp Arg Leu Lys Thr Pro Glu Glu His Glu Leu Glu Thr Gly 440 445 450 Ile Lys Ser Lys Glu Ala Arg Lys Tyr Ile Phe Asn Cys Leu Asp 455 460 465 Asp Met Ala Gln Val Asn Met Ser Thr Asp Leu Glu Gly Thr Asp 470 475 480 Met Leu Ala Glu Lys Ala Asp Arg Arg Glu Tyr Ile Asp Leu Leu 485 490 495 Lys Lys Met Leu Thr Ile Asp Ala Asp Lys Arg Ile Thr Pro Leu 500 505 510 Lys Thr Leu Asn His Gln Phe Val Thr Met Thr His Leu Leu Asp 515 520 525 Phe Pro His Ser Asn His Val Lys Ser Cys Phe Gln Asn Met Glu 530 535 540 Ile Cys Lys Arg Arg Val His Met Tyr Asp Thr Val Ser Gln Ile 545 550 555 Lys Ser Pro Phe Thr Thr His Val Ala Pro Asn Thr Ser Thr Asn 560 565 570 Leu Thr Met Ser Phe Ser Asn Gln Leu Asn Thr Val His Asn Gln 575 580 585 Ala Ser Val Leu Ala Ser Ser Ser Thr Ala Ala Ala Ala Thr Leu 590 595 600 Ser Leu Ala Asn Ser Asp Val Ser Leu Leu Asn Tyr Gln Ser Ala 605 610 615 Leu Tyr Pro Ser Ser Ala Ala Pro Val Pro Gly Val Ala Gln Gln 620 625 630 Gly Val Ser Leu Gln Pro Gly Thr Thr Gln Ile Cys Thr Gln Thr 635 640 645 Asp Pro Phe Gln Gln Thr Phe Ile Val Cys Pro Pro Ala Phe Gln 650 655 660 Thr Gly Leu Gln Ala Thr Thr Lys His Ser Gly Phe Pro Val Arg 665 670 675 Met Asp Asn Ala Val Pro Ile Val Pro Gln Ala Pro Ala Ala Gln 680 685 690 Pro Leu Gln Ile Gln Ser Gly Val Leu Thr Gln Gly Ser Cys Thr 695 700 705 Pro Leu Met Val Ala Thr Leu His Pro Gln Val Ala Thr Ile Thr 710 715 720 Pro Gln Tyr Ala Val Pro Phe Thr Leu Ser Cys Ala Ala Gly Arg 725 730 735 Pro Ala Leu Val Glu Gln Thr Ala Ala Val Leu Gln Ala Trp Pro 740 745 750 Gly Gly Thr Gln Gln Ile Leu Leu Pro Ser Thr Trp Gln Gln Leu 755 760 765 Pro Gly Val Ala Leu His Asn Ser Val Gln Pro Thr Ala Met Ile 770 775 780 Pro Glu Ala Met Gly Ser Gly Gln Gln Leu Ala Asp Trp Arg Asn 785 790 795 Ala His Ser His Gly Asn Gln Tyr Ser Thr Ile Met Gln Gln Pro 800 805 810 Ser Leu Leu Thr Asn His Val Thr Leu Ala Thr Ala Gln Pro Leu 815 820 825 Asn Val Gly Val Ala His Val Val Arg Gln Gln Gln Ser Ser Ser 830 835 840 Leu Pro Ser Lys Lys Asn Lys Gln Ser Ala Pro Val Ser Ser Lys 845 850 855 Ser Ser Leu Asp Val Leu Pro Ser Gln Val Tyr Ser Leu Val Gly 860 865 870 Ser Ser Pro Leu Arg Thr Thr Ser Ser Tyr Asn Ser Leu Val Pro 875 880 885 Val Gln Asp Gln His Gln Pro Ile Ile Ile Pro Asp Thr Pro Ser 890 895 900 Pro Pro Val Ser Val Ile Thr Ile Arg Ser Asp Thr Asp Glu Glu 905 910 915 Glu Asp Asn Lys Tyr Lys Pro Ser Ser Ser Gly Leu Lys Pro Arg 920 925 930 Ser Asn Val Ile Ser Tyr Val Thr Val Asn Asp Ser Pro Asp Ser 935 940 945 Asp Ser Ser Leu Ser Ser Pro Tyr Ser Thr Asp Thr Leu Ser Ala 950 955 960 Leu Arg Gly Asn Ser Gly Ser Val Leu Glu Gly Pro Gly Arg Val 965 970 975 Val Ala Asp Gly Thr Gly Thr Arg Thr Ile Ile Val Pro Pro Leu 980 985 990 Lys Thr Gln Leu Gly Asp Cys Thr Val Ala Thr Gln Ala Ser Gly 995 1000 1005 Leu Leu Ser Asn Lys Thr Lys Pro Val Ala Ser Val Ser Gly Gln 1010 1015 1020 Ser Ser Gly Cys Cys Ile Thr Pro Thr Gly Tyr Arg Ala Gln Arg 1025 1030 1035 Gly Gly Thr Ser Ala Ala Gln Pro Leu Asn Leu Ser Gln Asn Gln 1040 1045 1050 Gln Ser Ser Ala Ala Pro Thr Ser Gln Glu Arg Ser Ser Asn Pro 1055 1060 1065 Ala Pro Arg Arg Gln Gln Ala Phe Val Ala Pro Leu Ser Gln Ala 1070 1075 1080 Pro Tyr Thr Phe Gln His Gly Ser Pro Leu His Ser Thr Gly His 1085 1090 1095 Pro His Leu Ala Pro Ala Pro Ala His Leu Pro Ser Gln Ala His 1100 1105 1110 Leu Tyr Thr Tyr Ala Ala Pro Thr Ser Ala Ala Ala Leu Gly Ser 1115 1120 1125 Thr Ser Ser Ile Ala His Leu Phe Ser Pro Gln Gly Ser Ser Arg 1130 1135 1140 His Ala Ala Ala Tyr Thr Thr His Pro Ser Thr Leu Val His Gln 1145 1150 1155 Val Pro Val Ser Val Gly Pro Ser Leu Leu Thr Ser Ala Ser Val 1160 1165 1170 Ala Pro Ala Gln Tyr Gln His Gln Phe Ala Thr Gln Ser Tyr Ile 1175 1180 1185 Gly Ser Ser Arg Gly Ser Thr Ile Tyr Thr Gly Tyr Pro Leu Ser 1190 1195 1200 Pro Thr Lys Ile Ser Gln Tyr Ser Tyr Leu 1205 1210 2 357 PRT Homo sapiens misc_feature Incyte ID No 7472693CD1 2 Met Ala Arg Glu Asn Gly Glu Ser Ser Ser Ser Trp Lys Lys Gln 1 5 10 15 Ala Glu Asp Ile Lys Lys Ile Phe Glu Phe Lys Glu Thr Leu Gly 20 25 30 Thr Gly Ala Phe Ser Glu Val Val Leu Ala Glu Glu Lys Ala Thr 35 40 45 Gly Lys Leu Phe Ala Val Lys Cys Ile Pro Lys Lys Ala Leu Lys 50 55 60 Gly Lys Glu Ser Ser Ile Glu Asn Glu Ile Ala Val Leu Arg Lys 65 70 75 Ile Lys His Glu Asn Ile Val Ala Leu Glu Asp Ile Tyr Glu Ser 80 85 90 Pro Asn His Leu Tyr Leu Val Met Gln Leu Val Ser Gly Gly Glu 95 100 105 Leu Phe Asp Arg Ile Val Glu Lys Gly Phe Tyr Thr Glu Lys Asp 110 115 120 Ala Ser Thr Leu Ile Arg Gln Val Leu Asp Ala Val Tyr Tyr Leu 125 130 135 His Arg Met Gly Ile Val His Arg Asp Leu Lys Pro Glu Asn Leu 140 145 150 Leu Tyr Tyr Ser Gln Asp Glu Glu Ser Lys Ile Met Ile Ser Asp 155 160 165 Phe Gly Leu Ser Lys Met Glu Gly Lys Gly Asp Val Met Ser Thr 170 175 180 Ala Cys Gly Thr Pro Gly Tyr Val Ala Pro Glu Val Leu Ala Gln 185 190 195 Lys Pro Tyr Ser Lys Ala Val Asp Cys Trp Ser Ile Gly Val Ile 200 205 210 Ala Tyr Ile Leu Leu Cys Gly Tyr Pro Pro Phe Tyr Asp Glu Asn 215 220 225 Asp Ser Lys Leu Phe Glu Gln Ile Leu Lys Ala Glu Tyr Glu Phe 230 235 240 Asp Ser Pro Tyr Trp Asp Asp Ile Ser Asp Ser Ala Lys Asp Phe 245 250 255 Ile Arg Asn Leu Met Glu Lys Asp Pro Asn Lys Arg Tyr Thr Cys 260 265 270 Glu Gln Ala Ala Arg His Pro Trp Ile Ala Gly Asp Thr Ala Leu 275 280 285 Asn Lys Asn Ile His Glu Ser Val Ser Ala Gln Ile Arg Lys Asn 290 295 300 Phe Ala Lys Ser Lys Trp Arg Gln Ala Phe Asn Ala Thr Ala Val 305 310 315 Val Arg His Met Arg Lys Leu His Leu Gly Ser Ser Leu Asp Ser 320 325 330 Ser Asn Ala Ser Val Ser Ser Ser Leu Ser Leu Ala Ser Gln Lys 335 340 345 Asp Cys Ala Tyr Val Ala Lys Pro Glu Ser Leu Ser 350 355 3 419 PRT Homo sapiens misc_feature Incyte ID No 3107952CD1 3 Met Ala Gly Ser Gly Cys Ala Trp Gly Ala Glu Pro Pro Arg Phe 1 5 10 15 Leu Glu Ala Phe Gly Arg Leu Trp Gln Val Gln Ser Arg Leu Gly 20 25 30 Ser Gly Ser Ser Ala Ser Val Tyr Arg Val Arg Cys Cys Gly Asn 35 40 45 Pro Gly Ser Pro Pro Gly Ala Leu Lys Gln Phe Leu Pro Pro Gly 50 55 60 Thr Thr Gly Ala Ala Ala Ser Ala Ala Glu Tyr Gly Phe Arg Lys 65 70 75 Glu Arg Ala Ala Leu Glu Gln Leu Gln Gly His Arg Asn Ile Val 80 85 90 Thr Leu Tyr Gly Val Phe Thr Ile His Phe Ser Pro Asn Val Pro 95 100 105 Ser Arg Cys Leu Leu Leu Glu Leu Leu Asp Val Ser Val Ser Glu 110 115 120 Leu Leu Leu Tyr Ser Ser His Gln Gly Cys Ser Met Trp Met Ile 125 130 135 Gln His Cys Ala Arg Asp Val Leu Glu Ala Leu Ala Phe Leu His 140 145 150 His Glu Gly Tyr Val His Ala Asp Leu Lys Pro Arg Asn Ile Leu 155 160 165 Trp Ser Ala Glu Asn Glu Cys Phe Lys Leu Ile Asp Phe Gly Leu 170 175 180 Ser Phe Lys Glu Gly Asn Gln Asp Val Lys Tyr Ile Gln Thr Asp 185 190 195 Gly Tyr Arg Ala Pro Glu Ala Glu Leu Gln Asn Cys Leu Ala Gln 200 205 210 Ala Gly Leu Gln Ser Asp Thr Glu Cys Thr Ser Ala Val Asp Leu 215 220 225 Trp Ser Leu Gly Ile Ile Leu Leu Glu Met Phe Ser Gly Met Lys 230 235 240 Leu Lys His Thr Val Arg Ser Gln Glu Trp Lys Ala Asn Ser Ser 245 250 255 Ala Ile Ile Asp His Ile Phe Ala Ser Lys Ala Val Val Asn Ala 260 265 270 Ala Ile Pro Ala Tyr His Leu Arg Asp Leu Ile Lys Ser Met Leu 275 280 285 His Asp Asp Pro Ser Arg Arg Ile Pro Ala Glu Met Ala Leu Cys 290 295 300 Ser Pro Phe Phe Ser Ile Pro Phe Ala Pro His Ile Glu Asp Leu 305 310 315 Val Met Leu Pro Thr Pro Val Leu Arg Leu Leu Asn Val Leu Asp 320 325 330 Asp Asp Tyr Leu Glu Asn Glu Glu Glu Tyr Glu Asp Val Val Glu 335 340 345 Asp Val Lys Glu Glu Cys Gln Lys Tyr Gly Pro Val Val Ser Leu 350 355 360 Leu Val Pro Lys Gly Asn Pro Gly Arg Gly Gln Val Phe Val Glu 365 370 375 Tyr Ala Asn Ala Gly Asp Ser Lys Ala Ala Gln Lys Leu Leu Thr 380 385 390 Gly Arg Met Phe Asp Gly Lys Phe Val Val Ala Thr Phe Tyr Pro 395 400 405 Leu Ser Ala Tyr Lys Arg Gly Tyr Leu Tyr Gln Thr Leu Leu 410 415 4 624 PRT Homo sapiens misc_feature Incyte ID No 5544420CD1 4 Met Asn Arg Tyr Thr Thr Met Arg Gln Leu Gly Asp Gly Thr Tyr 1 5 10 15 Gly Ser Val Leu Met Gly Lys Ser Asn Glu Ser Gly Glu Leu Val 20 25 30 Ala Ile Lys Arg Met Lys Arg Lys Phe Tyr Ser Trp Asp Glu Cys 35 40 45 Met Asn Leu Arg Glu Val Lys Ser Leu Lys Lys Leu Asn His Ala 50 55 60 Asn Val Ile Lys Leu Lys Glu Val Ile Arg Glu Asn Asp His Leu 65 70 75 Tyr Phe Ile Phe Glu Tyr Met Lys Glu Asn Leu Tyr Gln Leu Met 80 85 90 Lys Asp Arg Asn Lys Leu Phe Pro Glu Ser Val Ile Arg Asn Ile 95 100 105 Met Tyr Gln Ile Leu Gln Gly Leu Ala Phe Ile His Lys His Gly 110 115 120 Phe Phe His Arg Asp Met Lys Pro Glu Asn Leu Leu Cys Met Gly 125 130 135 Pro Glu Leu Val Lys Ile Ala Asp Phe Gly Leu Ala Arg Glu Leu 140 145 150 Arg Ser Gln Pro Pro Tyr Thr Asp Tyr Val Ser Thr Arg Trp Tyr 155 160 165 Arg Ala Pro Glu Val Leu Leu Arg Ser Ser Val Tyr Ser Ser Pro 170 175 180 Ile Asp Val Trp Ala Val Gly Ser Ile Met Ala Glu Leu Tyr Met 185 190 195 Leu Arg Pro Leu Phe Pro Gly Thr Ser Glu Val Asp Glu Ile Phe 200 205 210 Lys Ile Cys Gln Val Leu Gly Thr Pro Lys Lys Ser Asp Trp Pro 215 220 225 Glu Gly Tyr Gln Leu Ala Ser Ser Met Asn Phe Arg Phe Pro Gln 230 235 240 Cys Val Pro Ile Asn Leu Lys Thr Leu Ile Pro Asn Ala Ser Asn 245 250 255 Glu Ala Ile Gln Leu Met Thr Glu Met Leu Asn Trp Asp Pro Lys 260 265 270 Lys Arg Pro Thr Ala Ser Gln Ala Leu Lys His Pro Tyr Phe Gln 275 280

285 Val Gly Gln Val Leu Gly Pro Ser Ser Asn His Leu Glu Ser Lys 290 295 300 Gln Ser Leu Asn Lys Gln Leu Gln Pro Leu Glu Ser Lys Pro Ser 305 310 315 Leu Val Glu Val Glu Pro Lys Pro Leu Pro Asp Ile Ile Asp Gln 320 325 330 Val Val Gly Gln Pro Gln Pro Lys Thr Ser Gln Gln Pro Leu Gln 335 340 345 Pro Ile Gln Pro Pro Gln Asn Leu Ser Val Gln Gln Pro Pro Lys 350 355 360 Gln Gln Ser Gln Glu Lys Pro Pro Gln Thr Leu Phe Pro Ser Ile 365 370 375 Val Lys Asn Met Pro Thr Lys Pro Asn Gly Thr Leu Ser His Lys 380 385 390 Ser Gly Arg Arg Arg Trp Gly Gln Thr Ile Phe Lys Ser Gly Asp 395 400 405 Ser Trp Glu Glu Leu Glu Asp Tyr Asp Phe Gly Ala Ser His Ser 410 415 420 Lys Lys Pro Ser Met Gly Val Phe Lys Glu Lys Arg Lys Lys Asp 425 430 435 Ser Pro Phe Arg Leu Pro Glu Pro Val Pro Ser Gly Ser Asn His 440 445 450 Ser Thr Gly Glu Asn Lys Ser Leu Pro Ala Val Thr Ser Leu Lys 455 460 465 Ser Asp Ser Glu Leu Ser Thr Ala Pro Thr Ser Lys Gln Tyr Tyr 470 475 480 Leu Lys Gln Ser Arg Tyr Leu Pro Gly Val Asn Pro Lys Lys Val 485 490 495 Ser Leu Ile Ala Ser Gly Lys Glu Ile Asn Pro His Thr Trp Ser 500 505 510 Asn Gln Leu Phe Pro Lys Ser Leu Gly Pro Val Gly Ala Glu Leu 515 520 525 Ala Phe Lys Arg Ser Asn Ala Glu Glu Lys Leu Gly Ser Tyr Ala 530 535 540 Thr Tyr Asn Gln Ser Gly Tyr Ile Pro Ser Phe Leu Lys Lys Glu 545 550 555 Val Gln Ser Ala Gly Gln Arg Ile His Leu Ala Pro Leu Asn Ala 560 565 570 Thr Ala Ser Glu Tyr Thr Trp Asn Thr Lys Thr Gly Arg Gly Gln 575 580 585 Phe Ser Gly Arg Thr Tyr Asn Pro Thr Ala Lys Asn Leu Asn Ile 590 595 600 Val Asn Arg Ala Gln Pro Ile Pro Ser Val His Gly Arg Thr Asp 605 610 615 Trp Val Ala Lys Tyr Gly Gly His Arg 620 5 878 PRT Homo sapiens misc_feature Incyte ID No 7472832CD1 5 Met Ala Thr Ala Pro Ser Tyr Pro Ala Gly Leu Pro Gly Ser Pro 1 5 10 15 Gly Pro Gly Ser Pro Pro Pro Pro Gly Gly Leu Glu Leu Gln Ser 20 25 30 Pro Pro Pro Leu Leu Pro Gln Ile Pro Ala Pro Gly Ser Gly Val 35 40 45 Ser Phe His Ile Gln Ile Gly Leu Thr Arg Glu Phe Val Leu Leu 50 55 60 Pro Ala Ala Ser Glu Leu Ala His Val Lys Gln Leu Ala Cys Ser 65 70 75 Ile Val Asp Gln Lys Phe Pro Glu Cys Gly Phe Tyr Gly Leu Tyr 80 85 90 Asp Lys Ile Leu Leu Phe Lys His Asp Pro Thr Ser Ala Asn Leu 95 100 105 Leu Gln Leu Val Arg Ser Ser Gly Asp Ile Gln Glu Gly Asp Leu 110 115 120 Val Glu Val Val Leu Ser Ala Ser Ala Thr Phe Glu Asp Phe Gln 125 130 135 Ile Arg Pro His Ala Leu Thr Val His Ser Tyr Arg Ala Pro Ala 140 145 150 Phe Cys Asp His Cys Gly Glu Met Leu Phe Gly Leu Val Arg Gln 155 160 165 Gly Leu Lys Cys Asp Gly Cys Gly Leu Asn Tyr His Lys Arg Cys 170 175 180 Ala Phe Ser Ile Pro Asn Asn Cys Ser Gly Ala Arg Lys Arg Arg 185 190 195 Leu Ser Ser Thr Ser Leu Ala Ser Gly His Ser Val Arg Leu Gly 200 205 210 Thr Ser Glu Ser Leu Pro Cys Thr Ala Glu Glu Leu Ser Arg Ser 215 220 225 Thr Thr Glu Leu Leu Pro Arg Arg Pro Pro Ser Ser Ser Ser Ser 230 235 240 Ser Ser Ala Ser Ser Tyr Thr Gly Arg Pro Ile Glu Leu Asp Lys 245 250 255 Met Leu Leu Ser Lys Val Lys Val Pro His Thr Phe Leu Ile His 260 265 270 Ser Tyr Thr Arg Pro Thr Val Cys Gln Ala Cys Lys Lys Leu Leu 275 280 285 Lys Gly Leu Phe Arg Gln Gly Leu Gln Cys Lys Asp Cys Lys Phe 290 295 300 Asn Cys His Lys Arg Cys Ala Thr Arg Val Pro Asn Asp Cys Leu 305 310 315 Gly Glu Ala Leu Ile Asn Gly Asp Val Pro Met Glu Glu Ala Thr 320 325 330 Asp Phe Ser Glu Ala Asp Lys Ser Ala Leu Met Asp Glu Ser Glu 335 340 345 Asp Ser Gly Val Ile Pro Gly Ser His Ser Glu Asn Ala Leu His 350 355 360 Ala Ser Glu Glu Glu Glu Gly Glu Gly Gly Lys Ala Gln Ser Ser 365 370 375 Leu Gly Tyr Ile Pro Leu Met Arg Val Val Gln Ser Val Arg His 380 385 390 Thr Thr Arg Lys Ser Ser Thr Thr Leu Arg Glu Gly Trp Val Val 395 400 405 His Tyr Ser Asn Lys Asp Thr Leu Arg Lys Arg His Tyr Trp Arg 410 415 420 Leu Asp Cys Lys Cys Ile Thr Leu Phe Gln Asn Asn Thr Thr Asn 425 430 435 Arg Tyr Tyr Lys Glu Ile Pro Leu Ser Glu Ile Leu Thr Val Glu 440 445 450 Ser Ala Gln Asn Phe Ser Leu Val Pro Pro Gly Thr Asn Pro His 455 460 465 Cys Phe Glu Ile Val Thr Ala Asn Ala Thr Tyr Phe Val Gly Glu 470 475 480 Met Pro Gly Gly Thr Pro Gly Gly Pro Ser Gly Gln Gly Ala Glu 485 490 495 Ala Ala Arg Gly Trp Glu Thr Ala Ile Arg Gln Ala Leu Met Pro 500 505 510 Val Ile Leu Gln Asp Ala Pro Ser Ala Pro Gly His Ala Pro His 515 520 525 Arg Gln Ala Ser Leu Ser Ile Ser Val Ser Asn Ser Gln Ile Gln 530 535 540 Glu Asn Val Asp Ile Ala Thr Val Tyr Gln Ile Phe Pro Asp Glu 545 550 555 Val Leu Gly Ser Gly Gln Phe Gly Val Val Tyr Gly Gly Lys His 560 565 570 Arg Lys Thr Gly Arg Asp Val Ala Val Lys Val Ile Asp Lys Leu 575 580 585 Arg Phe Pro Thr Lys Gln Glu Ser Gln Leu Arg Asn Glu Val Ala 590 595 600 Ile Leu Gln Ser Leu Arg His Pro Gly Ile Val Asn Leu Glu Cys 605 610 615 Met Phe Glu Thr Pro Glu Lys Val Phe Val Val Met Glu Lys Leu 620 625 630 His Gly Asp Met Leu Glu Met Ile Leu Ser Ser Glu Lys Gly Arg 635 640 645 Leu Pro Glu Arg Leu Thr Lys Phe Leu Ile Thr Gln Ile Leu Val 650 655 660 Ala Leu Arg His Leu His Phe Lys Asn Ile Val His Cys Asp Leu 665 670 675 Lys Pro Glu Asn Val Leu Leu Ala Ser Ala Asp Pro Phe Pro Gln 680 685 690 Val Lys Leu Cys Asp Phe Gly Phe Ala Arg Ile Ile Gly Glu Lys 695 700 705 Ser Phe Arg Arg Ser Val Val Gly Thr Pro Ala Tyr Leu Ala Pro 710 715 720 Glu Val Leu Leu Asn Gln Gly Tyr Asn Arg Ser Leu Asp Met Trp 725 730 735 Ser Val Gly Val Ile Met Tyr Val Ser Leu Ser Gly Thr Phe Pro 740 745 750 Phe Asn Glu Asp Glu Asp Ile Asn Asp Gln Ile Gln Asn Ala Ala 755 760 765 Phe Met Tyr Pro Ala Ser Pro Trp Ser His Ile Ser Ala Gly Ala 770 775 780 Ile Asp Leu Ile Asn Asn Leu Leu Gln Val Lys Met Arg Lys Arg 785 790 795 Tyr Ser Val Asp Lys Ser Leu Ser His Pro Trp Leu Gln Glu Tyr 800 805 810 Gln Thr Trp Leu Asp Leu Arg Glu Leu Glu Gly Lys Met Gly Glu 815 820 825 Arg Tyr Ile Thr His Glu Ser Asp Asp Ala Arg Trp Glu Gln Phe 830 835 840 Ala Ala Glu His Pro Leu Pro Gly Ser Gly Leu Pro Thr Asp Arg 845 850 855 Asp Leu Gly Gly Ala Cys Pro Pro Gln Asp His Asp Met Gln Gly 860 865 870 Leu Ala Glu Arg Ile Ser Val Leu 875 6 440 PRT Homo sapiens misc_feature Incyte ID No 1551456CD1 6 Met Ser Lys Leu Arg Met Lys Arg Arg Ala Ser Asp Arg Gly Ala 1 5 10 15 Gly Glu Thr Ser Ala Arg Ala Lys Ala Leu Gly Ser Gly Ile Ser 20 25 30 Gly Asn Asn Ala Lys Arg Ala Gly Pro Phe Ile Leu Gly Pro Arg 35 40 45 Leu Gly Asn Ser Pro Val Pro Ser Ile Val Gln Cys Leu Ala Arg 50 55 60 Lys Asp Gly Thr Asp Asp Phe Tyr Gln Leu Lys Ile Leu Thr Leu 65 70 75 Glu Glu Arg Gly Asp Gln Gly Ile Glu Ser Gln Glu Glu Arg Gln 80 85 90 Gly Lys Met Leu Leu His Thr Glu Tyr Ser Leu Leu Ser Leu Leu 95 100 105 His Thr Gln Asp Gly Val Val His His His Gly Leu Phe Gln Asp 110 115 120 Arg Thr Cys Glu Ile Val Glu Asp Thr Glu Ser Ser Arg Met Val 125 130 135 Lys Lys Met Lys Lys Arg Ile Cys Leu Val Leu Asp Cys Leu Cys 140 145 150 Ala His Asp Phe Ser Asp Lys Thr Ala Asp Leu Ile Asn Leu Gln 155 160 165 His Tyr Val Ile Lys Glu Lys Arg Leu Ser Glu Arg Glu Thr Val 170 175 180 Val Ile Phe Tyr Asp Val Val Arg Val Val Glu Ala Leu His Gln 185 190 195 Lys Asn Ile Val His Arg Asp Leu Lys Leu Gly Asn Met Val Leu 200 205 210 Asn Lys Arg Thr His Arg Ile Thr Ile Thr Asn Phe Cys Leu Gly 215 220 225 Lys His Leu Val Ser Glu Gly Asp Leu Leu Lys Asp Gln Arg Gly 230 235 240 Ser Pro Ala Tyr Ile Ser Pro Asp Val Leu Ser Gly Arg Pro Tyr 245 250 255 Arg Gly Lys Pro Ser Asp Met Trp Ala Leu Gly Val Val Leu Phe 260 265 270 Thr Met Leu Tyr Gly Gln Phe Pro Phe Tyr Asp Ser Ile Pro Gln 275 280 285 Glu Leu Phe Arg Lys Ile Lys Ala Ala Glu Tyr Thr Ile Pro Glu 290 295 300 Asp Gly Arg Val Ser Glu Asn Thr Val Cys Leu Ile Arg Lys Leu 305 310 315 Leu Val Leu Asp Pro Gln Gln Arg Leu Ala Ala Ala Asp Val Leu 320 325 330 Glu Ala Leu Ser Ala Ile Ile Ala Ser Trp Gln Ser Leu Ser Ser 335 340 345 Leu Ser Gly Pro Leu Gln Val Val Pro Asp Ile Asp Asp Gln Met 350 355 360 Ser Asn Ala Asp Ser Ser Gln Glu Ala Lys Val Thr Glu Glu Cys 365 370 375 Ser Gln Tyr Glu Phe Glu Asn Tyr Met Arg Gln Gln Leu Leu Leu 380 385 390 Ala Glu Glu Lys Ser Ser Ile His Asp Ala Arg Ser Trp Val Pro 395 400 405 Lys Arg Gln Phe Gly Ser Ala Pro Pro Val Arg Arg Leu Gly His 410 415 420 Asp Ala Gln Pro Met Thr Ser Leu Asp Thr Ala Ile Leu Ala Gln 425 430 435 Arg Tyr Leu Arg Lys 440 7 923 PRT Homo sapiens misc_feature Incyte ID No 2589355CD1 7 Met Ala Arg Gly Thr Cys Ser Ala Gly Arg Ser Gly Trp Gly Ser 1 5 10 15 Thr Thr Ser Arg Ala Arg Trp Ala Ser Gly Asn Phe Ala Val Val 20 25 30 Lys Leu Gly Arg His Arg Ile Thr Lys Thr Glu Val Ala Ile Lys 35 40 45 Ile Ile Asp Lys Ser Gln Pro Trp Met His Val Asn Leu Glu Lys 50 55 60 Ile Tyr Arg Glu Val Gln Ile Met Lys Met Leu Asp His Pro His 65 70 75 Ile Ile Lys Leu Tyr Gln Val Met Glu Thr Lys Ser Met Leu Tyr 80 85 90 Leu Val Thr Glu Tyr Ala Lys Asn Gly Glu Ile Phe Asp Tyr Leu 95 100 105 Ala Asn His Gly Arg Leu Asn Glu Ser Glu Ala Arg Arg Lys Phe 110 115 120 Trp Gln Ile Leu Ser Ala Val Asp Tyr Cys His Gly Arg Lys Ile 125 130 135 Val His Arg Asp Leu Lys Ala Glu Asn Leu Leu Leu Asp Asn Asn 140 145 150 Met Asn Ile Lys Ile Ala Asp Phe Gly Phe Gly Asn Phe Phe Lys 155 160 165 Ser Gly Glu Leu Leu Ala Thr Trp Cys Gly Ser Pro Pro Tyr Ala 170 175 180 Ala Pro Glu Val Phe Glu Gly Gln Gln Tyr Glu Gly Pro Gln Leu 185 190 195 Asp Ile Trp Ser Met Gly Val Val Leu Tyr Val Leu Val Cys Gly 200 205 210 Ala Leu Pro Phe Asp Gly Pro Thr Leu Pro Ile Leu Arg Gln Arg 215 220 225 Val Leu Glu Gly Arg Phe Arg Ile Pro Tyr Phe Met Ser Glu Asp 230 235 240 Cys Glu His Leu Ile Arg Arg Met Leu Val Leu Asp Pro Ser Lys 245 250 255 Arg Leu Thr Ile Ala Gln Ile Lys Glu His Lys Trp Met Leu Ile 260 265 270 Glu Val Pro Val Gln Arg Pro Val Leu Tyr Pro Gln Glu Gln Glu 275 280 285 Asn Glu Pro Ser Ile Gly Glu Phe Asn Glu Gln Val Leu Arg Leu 290 295 300 Met His Ser Leu Gly Ile Asp Gln Gln Lys Thr Ile Glu Ser Leu 305 310 315 Gln Asn Lys Ser Tyr Asn His Phe Ala Ala Ile Tyr Phe Leu Leu 320 325 330 Val Glu Arg Leu Lys Ser His Arg Ser Ser Phe Pro Val Glu Gln 335 340 345 Arg Leu Asp Gly Arg Gln Arg Arg Pro Ser Thr Ile Ala Glu Gln 350 355 360 Thr Val Ala Lys Ala Gln Thr Val Gly Leu Pro Val Thr Met His 365 370 375 Ser Pro Asn Met Arg Leu Leu Arg Ser Ala Leu Leu Pro Gln Ala 380 385 390 Ser Asn Val Glu Ala Phe Ser Phe Pro Ala Ser Gly Cys Gln Ala 395 400 405 Glu Ala Ala Phe Met Glu Glu Glu Cys Val Asp Thr Pro Lys Val 410 415 420 Asn Gly Cys Leu Leu Asp Pro Val Pro Pro Val Leu Val Arg Lys 425 430 435 Gly Cys Gln Ser Leu Pro Ser Asn Met Met Glu Thr Ser Ile Asp 440 445 450 Glu Gly Leu Glu Thr Glu Gly Glu Ala Glu Glu Asp Pro Ala His 455 460 465 Ala Phe Glu Ala Phe Gln Ser Thr Arg Ser Gly Gln Arg Arg His 470 475 480 Thr Leu Ser Glu Val Thr Asn Gln Leu Val Val Met Pro Gly Ala 485 490 495 Gly Lys Ile Phe Ser Met Asn Asp Ser Pro Ser Leu Asp Ser Val 500 505 510 Asp Ser Glu Tyr Asp Met Gly Ser Val Gln Arg Asp Leu Asn Phe 515 520 525 Leu Glu Asp Asn Pro Ser Leu Lys Asp Ile Met Leu Ala Asn Gln 530 535 540 Pro Ser Pro Arg Met Thr Ser Pro Phe Ile Ser Leu Arg Pro Thr 545 550 555 Asn Pro Ala Met Gln Ala Leu Ser Ser Gln Lys Arg Glu Val His 560 565 570 Asn Arg Ser Pro Val Ser Phe Arg Glu Gly Arg Arg Ala Ser Asp 575 580 585 Thr Ser Leu Thr Gln Gly Ile Val Ala Phe Arg Gln His Leu Gln 590 595 600 Asn Leu Ala Arg Thr Lys Gly Ile Leu Glu Leu Asn Lys Val Gln 605 610 615 Leu Leu Tyr Glu Gln Ile Gly Pro Glu

Ala Asp Pro Asn Leu Ala 620 625 630 Pro Ala Ala Pro Gln Leu Gln Asp Leu Ala Ser Ser Cys Pro Gln 635 640 645 Glu Glu Val Ser Gln Gln Gln Glu Ser Val Ser Thr Leu Pro Ala 650 655 660 Ser Val His Pro Gln Leu Ser Pro Arg Gln Ser Leu Glu Thr Gln 665 670 675 Tyr Leu Gln His Arg Leu Gln Lys Pro Ser Leu Leu Ser Lys Ala 680 685 690 Gln Asn Thr Cys Gln Leu Tyr Cys Lys Glu Pro Pro Arg Ser Leu 695 700 705 Glu Gln Gln Leu Gln Glu His Arg Leu Gln Gln Lys Arg Leu Phe 710 715 720 Leu Gln Lys Gln Ser Gln Leu Gln Ala Tyr Phe Asn Gln Met Gln 725 730 735 Ile Ala Glu Ser Ser Tyr Pro Gln Pro Ser Gln Gln Leu Pro Leu 740 745 750 Pro Arg Gln Glu Thr Pro Pro Pro Ser Gln Gln Ala Pro Pro Phe 755 760 765 Ser Leu Thr Gln Pro Leu Ser Pro Val Leu Glu Pro Ser Ser Glu 770 775 780 Gln Met Gln Tyr Ser Pro Phe Leu Ser Gln Tyr Gln Glu Met Gln 785 790 795 Leu Gln Pro Leu Pro Ser Thr Ser Gly Pro Arg Ala Ala Pro Pro 800 805 810 Leu Pro Thr Gln Leu Gln Gln Gln Gln Pro Pro Pro Pro Pro Pro 815 820 825 Pro Pro Pro Pro Arg Gln Pro Gly Ala Ala Pro Ala Pro Leu Gln 830 835 840 Phe Ser Tyr Gln Thr Cys Glu Leu Pro Ser Ala Ala Ser Pro Ala 845 850 855 Pro Asp Tyr Pro Thr Pro Cys Gln Tyr Pro Val Asp Gly Ala Gln 860 865 870 Gln Ser Asp Leu Thr Gly Pro Asp Cys Pro Arg Ser Pro Gly Leu 875 880 885 Gln Glu Ala Pro Ser Ser Tyr Asp Pro Leu Ala Leu Ser Glu Leu 890 895 900 Pro Gly Leu Phe Asp Cys Glu Met Leu Asp Ala Val Asp Pro Gln 905 910 915 His Asn Gly Tyr Val Leu Val Asn 920 8 442 PRT Homo sapiens misc_feature Incyte ID No 4357117CD1 8 Met Arg Ile Val Cys Leu Val Lys Asn Gln Gln Pro Leu Gly Ala 1 5 10 15 Thr Ile Lys Arg His Glu Met Thr Gly Asp Ile Leu Val Ala Arg 20 25 30 Ile Ile His Gly Gly Leu Ala Glu Arg Ser Gly Leu Leu Tyr Ala 35 40 45 Gly Asp Lys Leu Val Glu Val Asn Gly Val Ser Val Glu Gly Leu 50 55 60 Asp Pro Glu Gln Val Ile His Ile Leu Ala Met Ser Arg Gly Thr 65 70 75 Ile Met Phe Lys Val Val Pro Val Ser Asp Pro Pro Val Asn Ser 80 85 90 Gln Gln Met Val Tyr Val Arg Ala Met Thr Glu Tyr Trp Pro Gln 95 100 105 Glu Asp Pro Asp Ile Pro Cys Met Asp Ala Gly Leu Pro Phe Gln 110 115 120 Lys Gly Asp Ile Leu Gln Ile Val Asp Gln Asn Asp Ala Leu Trp 125 130 135 Trp Gln Ala Arg Lys Ile Ser Asp Pro Ala Thr Cys Ala Gly Leu 140 145 150 Val Pro Ser Asn His Leu Leu Lys Arg Lys Gln Arg Glu Phe Trp 155 160 165 Trp Ser Gln Pro Tyr Gln Pro His Thr Cys Leu Lys Ser Thr Ser 170 175 180 Asp Lys Glu Glu Phe Val Gly Tyr Gly Gln Lys Phe Phe Ile Gly 185 190 195 Arg Phe Ser Pro Leu His Ala Ser Val Cys Cys Thr Gly Ser Cys 200 205 210 Tyr Ser Ala Val Gly Ala Pro Tyr Glu Glu Val Val Arg Tyr Gln 215 220 225 Arg Arg Pro Ser Asp Lys Tyr Arg Leu Ile Val Leu Ile Gly Pro 230 235 240 Ser Gly Val Gly Val Asn Glu Leu Arg Arg Gln Leu Ile Glu Phe 245 250 255 Asn Pro Ser His Phe Gln Ser Ala Val Pro His Thr Thr Arg Thr 260 265 270 Lys Lys Ser Tyr Glu Met Asn Gly Arg Glu Tyr His Tyr Val Ser 275 280 285 Lys Glu Thr Phe Glu Asn Leu Ile Tyr Ser His Arg Met Leu Glu 290 295 300 Tyr Gly Glu Tyr Lys Gly His Leu Tyr Gly Thr Ser Val Asp Ala 305 310 315 Val Gln Thr Val Leu Val Glu Gly Lys Ile Cys Val Met Asp Leu 320 325 330 Glu Pro Gln Asp Ile Gln Gly Val Arg Thr His Glu Leu Lys Pro 335 340 345 Tyr Val Ile Phe Ile Lys Pro Ser Asn Met Arg Cys Met Lys Gln 350 355 360 Ser Arg Lys Asn Ala Lys Val Ile Thr Asp Tyr Tyr Val Asp Met 365 370 375 Lys Phe Lys Asp Glu Asp Leu Gln Glu Met Glu Asn Leu Ala Gln 380 385 390 Arg Met Glu Thr Gln Phe Gly Gln Phe Phe Asp His Val Ile Val 395 400 405 Asn Asp Ser Leu His Asp Ala Cys Ala Gln Leu Leu Ser Ala Ile 410 415 420 Gln Lys Ala Gln Glu Glu Pro Gln Trp Val Pro Ala Thr Trp Ile 425 430 435 Ser Ser Asp Thr Glu Ser Gln 440 9 1046 PRT Homo sapiens misc_feature Incyte ID No 5511992CD1 9 Met Glu Pro Ser Arg Ala Leu Leu Gly Cys Leu Ala Ser Ala Ala 1 5 10 15 Ala Ala Ala Pro Pro Gly Glu Asp Gly Ala Gly Ala Gly Ala Glu 20 25 30 Glu Glu Glu Glu Glu Glu Glu Glu Ala Ala Ala Ala Val Gly Pro 35 40 45 Gly Glu Leu Gly Cys Asp Ala Pro Leu Pro Tyr Trp Thr Ala Val 50 55 60 Phe Glu Tyr Glu Ala Ala Gly Glu Asp Glu Leu Thr Leu Arg Leu 65 70 75 Gly Asp Val Val Glu Val Leu Ser Lys Asp Ser Gln Val Ser Gly 80 85 90 Asp Glu Gly Trp Trp Thr Gly Gln Leu Asn Gln Arg Val Gly Ile 95 100 105 Phe Pro Ser Asn Tyr Val Thr Pro Arg Ser Ala Phe Ser Ser Arg 110 115 120 Cys Gln Pro Gly Gly Glu Glu Glu Ile Asp Phe Ala Glu Leu Thr 125 130 135 Leu Glu Glu Ile Ile Gly Ile Gly Gly Phe Gly Lys Val Tyr Arg 140 145 150 Ala Phe Trp Ile Gly Asp Glu Val Ala Val Lys Ala Ala Arg His 155 160 165 Asp Pro Asp Glu Asp Ile Ser Gln Thr Ile Glu Asn Val Arg Gln 170 175 180 Glu Ala Lys Leu Phe Ala Met Leu Lys His Pro Asn Ile Ile Ala 185 190 195 Leu Arg Gly Val Cys Leu Lys Glu Pro Asn Leu Cys Leu Val Met 200 205 210 Glu Phe Ala Arg Gly Gly Pro Leu Asn Arg Val Leu Ser Gly Lys 215 220 225 Arg Ile Pro Pro Asp Ile Leu Val Asn Trp Ala Val Gln Ile Ala 230 235 240 Arg Gly Met Asn Tyr Leu Leu Asp Glu Ala Ile Val Pro Ile Ile 245 250 255 His Arg Asp Leu Lys Ser Ser Asn Ile Leu Ile Leu Gln Lys Val 260 265 270 Glu Asn Gly Asp Leu Ser Asn Lys Ile Leu Lys Ile Thr Asp Phe 275 280 285 Gly Leu Ala Arg Glu Trp His Arg Thr Thr Lys Met Ser Ala Ala 290 295 300 Gly Thr Tyr Ala Trp Met Ala Pro Glu Val Ile Arg Ala Ser Met 305 310 315 Phe Ser Lys Gly Ser Asp Val Trp Ser Tyr Gly Val Leu Leu Trp 320 325 330 Glu Leu Leu Thr Gly Glu Val Pro Phe Arg Gly Ile Asp Gly Leu 335 340 345 Ala Val Ala Tyr Gly Val Ala Met Asn Lys Leu Ala Leu Pro Ile 350 355 360 Pro Ser Thr Cys Pro Glu Pro Phe Ala Lys Leu Met Glu Asp Cys 365 370 375 Trp Asn Pro Asp Pro His Ser Arg Pro Ser Phe Thr Asn Ile Leu 380 385 390 Asp Gln Leu Thr Thr Ile Glu Glu Ser Gly Phe Phe Glu Met Pro 395 400 405 Lys Asp Ser Phe His Cys Leu Gln Asp Asn Trp Lys His Glu Ile 410 415 420 Gln Glu Met Phe Asp Gln Leu Arg Ala Lys Glu Lys Glu Leu Arg 425 430 435 Thr Trp Glu Glu Glu Leu Thr Arg Ala Ala Leu Gln Gln Lys Asn 440 445 450 Gln Glu Glu Leu Leu Arg Arg Arg Glu Gln Glu Leu Ala Glu Arg 455 460 465 Glu Ile Asp Ile Leu Glu Arg Glu Leu Asn Ile Ile Ile His Gln 470 475 480 Leu Cys Gln Glu Lys Pro Arg Val Lys Lys Arg Lys Gly Lys Phe 485 490 495 Arg Lys Ser Arg Leu Lys Leu Lys Asp Gly Asn Arg Ile Ser Leu 500 505 510 Pro Ser Gly Phe Gln His Lys Phe Thr Val Gln Ala Ser Pro Thr 515 520 525 Met Asp Lys Arg Lys Ser Leu Ile Asn Ser Arg Ser Ser Pro Pro 530 535 540 Ala Ser Pro Thr Ile Ile Pro Arg Leu Arg Ala Ile Gln Cys Glu 545 550 555 Thr Val Ser Lys Thr Trp Gly Arg Ser Ser Val Val Pro Lys Glu 560 565 570 Glu Gly Glu Glu Glu Glu Lys Arg Ala Pro Lys Lys Lys Gly Arg 575 580 585 Thr Trp Gly Pro Gly Thr Leu Gly Gln Lys Glu Leu Ala Ser Gly 590 595 600 Asp Glu Ser Leu Lys Ser Leu Val Asp Gly Tyr Lys Gln Trp Ser 605 610 615 Ser Ser Ala Pro Asn Leu Val Lys Gly Pro Arg Ser Ser Pro Ala 620 625 630 Leu Pro Gly Phe Thr Ser Leu Met Glu Met Gly Lys Phe Thr Glu 635 640 645 Asp Glu Asp Ser Glu Gly Pro Gly Ser Gly Glu Ser Arg Leu Gln 650 655 660 His Ser Pro Ser Gln Ser Tyr Leu Cys Ile Pro Phe Pro Arg Gly 665 670 675 Glu Asp Gly Asp Gly Pro Ser Ser Asp Gly Ile His Glu Glu Pro 680 685 690 Thr Pro Val Asn Ser Ala Thr Ser Thr Pro Gln Leu Thr Pro Thr 695 700 705 Asn Ser Leu Lys Arg Gly Gly Ala His His Arg Arg Cys Glu Val 710 715 720 Ala Leu Leu Gly Cys Gly Ala Val Leu Ala Ala Thr Gly Leu Gly 725 730 735 Phe Asp Leu Leu Glu Ala Gly Lys Cys Gln Leu Leu Pro Leu Glu 740 745 750 Glu Pro Glu Pro Pro Ala Arg Glu Glu Lys Lys Arg Arg Glu Gly 755 760 765 Leu Phe Gln Arg Ser Ser Arg Pro Arg Arg Ser Thr Ser Pro Pro 770 775 780 Ser Arg Lys Leu Phe Lys Lys Glu Glu Pro Met Leu Leu Leu Gly 785 790 795 Asp Pro Ser Ala Ser Leu Thr Leu Leu Ser Leu Ser Ser Ile Ser 800 805 810 Glu Cys Asn Ser Thr Arg Ser Leu Leu Arg Ser Asp Ser Asp Glu 815 820 825 Ile Val Val Tyr Glu Met Pro Val Ser Pro Val Glu Ala Pro Pro 830 835 840 Leu Ser Pro Cys Thr His Asn Pro Leu Val Asn Val Arg Val Glu 845 850 855 Arg Phe Lys Arg Asp Pro Asn Gln Ser Leu Thr Pro Thr His Val 860 865 870 Thr Leu Thr Thr Pro Ser Gln Pro Ser Ser His Arg Arg Thr Pro 875 880 885 Ser Asp Gly Ala Leu Pro Ser Pro Ser Arg Asp Pro Gly Glu Phe 890 895 900 Pro Arg Leu Pro Asp Pro Asn Val Val Phe Pro Pro Thr Pro Arg 905 910 915 Arg Trp Asn Thr Gln Gln Asp Ser Thr Leu Glu Arg Pro Lys Thr 920 925 930 Leu Glu Phe Leu Pro Arg Pro Arg Pro Ser Ala Asn Arg Gln Arg 935 940 945 Leu Asp Pro Trp Trp Phe Val Ser Pro Ser His Ala Arg Ser Thr 950 955 960 Ser Pro Ala Asn Ser Ser Ser Thr Glu Thr Pro Ser Asn Leu Asp 965 970 975 Ser Cys Phe Ala Ser Ser Ser Ser Thr Val Glu Glu Arg Pro Gly 980 985 990 Leu Pro Ala Leu Leu Pro Phe Gln Ala Gly Pro Leu Pro Pro Thr 995 1000 1005 Glu Arg Thr Leu Leu Asp Leu Asp Ala Glu Gly Gln Ser Gln Asp 1010 1015 1020 Ser Thr Val Pro Leu Cys Arg Ala Glu Leu Asn Thr His Arg Pro 1025 1030 1035 Ala Pro Tyr Glu Ile Gln Gln Glu Phe Trp Ser 1040 1045 10 357 PRT Homo sapiens misc_feature Incyte ID No 7474560CD1 10 Met Gln Ile Pro Asp Glu Glu Gly Ile Val Ile Asp Gly Phe Pro 1 5 10 15 Arg Asp Val Ala Gln Ala Leu Ser Phe Glu Asp Gln Ile Cys Thr 20 25 30 Pro Asp Leu Val Val Phe Leu Ala Cys Ala Asn Gln Arg Leu Lys 35 40 45 Glu Arg Leu Leu Lys Arg Ala Glu Gln Gln Gly Arg Pro Asp Asp 50 55 60 Asn Val Lys Ala Thr Gln Arg Arg Leu Met Asn Phe Lys Gln Asn 65 70 75 Ala Ala Pro Leu Val Lys Tyr Phe Gln Glu Lys Gly Leu Ile Met 80 85 90 Thr Phe Asp Ala Asp Arg Asp Glu Asp Glu Val Phe Tyr Asp Ile 95 100 105 Ser Met Ala Val Asp Asn Lys Leu Phe Pro Asn Lys Glu Ala Ala 110 115 120 Ala Gly Ser Ser Asp Leu Asp Pro Ser Met Ile Leu Asp Thr Gly 125 130 135 Glu Ile Ile Asp Thr Gly Ser Asp Tyr Glu Asp Gln Gly Asp Asp 140 145 150 Gln Leu Asn Val Phe Gly Glu Asp Thr Met Gly Gly Phe Met Glu 155 160 165 Asp Leu Arg Lys Cys Lys Ile Ile Phe Ile Ile Gly Gly Pro Gly 170 175 180 Ser Gly Lys Gly Thr Gln Cys Glu Lys Leu Val Glu Lys Tyr Gly 185 190 195 Phe Thr His Leu Ser Thr Gly Glu Leu Leu Arg Glu Glu Leu Ala 200 205 210 Ser Glu Ser Glu Arg Ser Lys Leu Ile Arg Asp Ile Met Glu Arg 215 220 225 Gly Asp Leu Val Pro Ser Gly Ile Val Leu Glu Leu Leu Lys Glu 230 235 240 Ala Met Val Ala Ser Leu Gly Asp Thr Arg Gly Phe Leu Ile Asp 245 250 255 Gly Tyr Pro Arg Glu Val Lys Gln Gly Glu Glu Phe Gly Arg Arg 260 265 270 Ile Gly Asp Pro Gln Leu Val Ile Cys Met Asp Cys Ser Ala Asp 275 280 285 Thr Met Thr Asn Arg Leu Leu Gln Arg Ser Arg Ser Ser Leu Pro 290 295 300 Val Asp Asp Thr Thr Lys Thr Ile Ala Lys Arg Leu Glu Ala Tyr 305 310 315 Tyr Arg Ala Ser Ile Pro Val Ile Ala Tyr Tyr Glu Thr Lys Thr 320 325 330 Gln Leu His Lys Ile Asn Ala Glu Gly Thr Pro Glu Asp Val Phe 335 340 345 Leu Gln Leu Cys Thr Ala Ile Asp Ser Ile Ile Phe 350 355 11 355 PRT Homo sapiens misc_feature Incyte ID No 7474602CD1 11 Met Ala Arg Glu Asn Gly Glu Ser Ser Ser Ser Trp Lys Lys Gln 1 5 10 15 Ala Glu Asp Ile Lys Lys Ile Phe Glu Phe Lys Glu Thr Leu Gly 20 25 30 Thr Gly Ala Phe Ser Glu Val Val Leu Ala Glu Glu Lys Ala Thr 35 40 45 Gly Lys Leu Phe Ala Val Lys Cys Ile Pro Lys Lys Ala Leu Lys 50 55 60 Gly Lys Glu Ser Ser Ile Glu Asn Glu Ile Ala Val Leu Arg Lys 65 70 75 Ile Lys His Glu Asn Ile Val Ala Leu Glu Asp Ile Tyr Glu Ser 80 85 90 Pro Asn His Leu Tyr Leu Val Met Gln Leu Val Ser Gly Gly Glu 95 100 105 Leu Phe Asp Arg Ile Val Glu Lys Gly Phe Tyr Thr Glu Lys Asp 110 115 120 Ala Ser Thr Leu Ile Arg Gln

Val Leu Asp Ala Val Tyr Tyr Leu 125 130 135 His Arg Met Gly Ile Val His Arg Asp Leu Lys Pro Glu Asn Leu 140 145 150 Leu Tyr Tyr Ser Gln Asp Glu Glu Ser Lys Ile Met Ile Ser Asp 155 160 165 Phe Gly Leu Ser Lys Met Glu Gly Lys Gly Asp Val Met Ser Thr 170 175 180 Ala Cys Gly Thr Pro Gly Tyr Val Ala Pro Glu Val Leu Ala Gln 185 190 195 Lys Pro Tyr Ser Lys Ala Val Asp Cys Trp Ser Ile Gly Val Ile 200 205 210 Ala Tyr Ile Leu Leu Cys Gly Tyr Pro Pro Phe Tyr Asp Glu Asn 215 220 225 Asp Ser Lys Leu Phe Glu Gln Ile Leu Lys Ala Glu Tyr Glu Phe 230 235 240 Asp Ser Pro Tyr Trp Asp Asp Ile Ser Asp Ser Ala Lys Asp Phe 245 250 255 Ile Arg Asn Leu Met Glu Lys Asp Pro Asn Lys Arg Tyr Thr Cys 260 265 270 Glu Gln Ala Ala Arg His Pro Trp Ile Ala Gly Asp Thr Ala Leu 275 280 285 Asn Lys Asn Ile His Glu Ser Val Ser Ala Gln Ile Arg Lys Asn 290 295 300 Phe Ala Lys Ser Lys Trp Arg Gln Ala Phe Asn Ala Thr Ala Val 305 310 315 Val Arg His Met Arg Lys Leu His Leu Gly Ser Ser Leu Asp Ser 320 325 330 Ser Asn Ala Ser Val Ser Ser Ser Leu Ser Leu Ala Ser Gln Lys 335 340 345 Asp Cys Ala Ser Gly Thr Phe His Ala Leu 350 355 12 224 PRT Homo sapiens misc_feature Incyte ID No 7475509CD1 12 Met Ala Ser Lys Leu Leu Arg Ala Val Ile Leu Gly Pro Pro Gly 1 5 10 15 Ser Gly Lys Gly Thr Val Cys Gln Arg Ile Ala Gln Asn Phe Gly 20 25 30 Leu Gln His Leu Ser Ser Gly His Phe Leu Arg Glu Asn Ile Lys 35 40 45 Ala Ser Thr Glu Val Gly Glu Met Ala Lys Gln Tyr Ile Glu Lys 50 55 60 Ser Leu Leu Val Pro Asp His Val Ile Thr Arg Leu Met Met Ser 65 70 75 Glu Leu Glu Asn Arg Arg Gly Gln His Trp Leu Leu Asp Gly Phe 80 85 90 Pro Arg Thr Leu Gly Gln Ala Glu Ala Leu Asp Lys Ile Cys Glu 95 100 105 Val Asp Leu Val Ile Ser Leu Asn Ile Pro Phe Glu Thr Leu Lys 110 115 120 Asp Arg Leu Ser Arg Arg Trp Ile His Pro Pro Ser Gly Arg Val 125 130 135 Tyr Asn Leu Asp Phe Asn Pro Pro His Val His Gly Ile Asp Asp 140 145 150 Val Thr Gly Glu Pro Leu Val Gln Gln Glu Asp Asp Lys Pro Glu 155 160 165 Ala Val Ala Ala Arg Leu Arg Gln Tyr Lys Asp Val Ala Lys Pro 170 175 180 Val Ile Glu Leu Tyr Lys Ser Arg Gly Val Leu His Gln Phe Phe 185 190 195 Arg Asn Arg Arg Arg Thr Lys Ile Trp Pro Tyr Val Tyr Thr Thr 200 205 210 Phe Leu Asn Lys Ile Thr Pro Ile Gln Ser Lys Glu Ala Phe 215 220 13 502 PRT Homo sapiens misc_feature Incyte ID No 7475491CD1 13 Met Asn Lys Met Lys Asn Phe Lys Arg Arg Phe Ser Leu Ser Val 1 5 10 15 Pro Arg Thr Glu Thr Ile Glu Glu Ser Leu Ala Glu Phe Thr Glu 20 25 30 Gln Phe Asn Gln Leu His Asn Arg Arg Asn Glu Asn Leu Gln Leu 35 40 45 Gly Pro Leu Gly Arg Asp Pro Pro Gln Glu Cys Ser Thr Phe Ser 50 55 60 Pro Thr Asp Ser Gly Glu Glu Pro Gly Gln Leu Ser Pro Gly Val 65 70 75 Gln Phe Gln Arg Arg Gln Asn Gln Arg Arg Phe Ser Met Glu Val 80 85 90 Arg Ala Ser Gly Ala Leu Pro Arg Gln Val Ala Gly Cys Thr His 95 100 105 Lys Gly Val His Arg Arg Ala Ala Ala Leu Gln Pro Asp Phe Asp 110 115 120 Val Ser Lys Arg Leu Ser Leu Pro Met Asp Ile Arg Leu Pro Gln 125 130 135 Glu Phe Leu Gln Lys Leu Gln Met Glu Ser Pro Asp Leu Pro Lys 140 145 150 Pro Leu Ser Arg Met Ser Arg Arg Ala Ser Leu Ser Asp Ile Gly 155 160 165 Phe Gly Lys Leu Glu Thr Tyr Val Lys Leu Asp Lys Leu Gly Glu 170 175 180 Gly Thr Tyr Ala Thr Val Phe Lys Gly Arg Ser Lys Leu Thr Glu 185 190 195 Asn Leu Val Ala Leu Lys Glu Ile Arg Leu Glu His Glu Glu Gly 200 205 210 Ala Pro Cys Thr Ala Ile Arg Glu Val Ser Leu Leu Lys Asn Leu 215 220 225 Lys His Ala Asn Ile Val Thr Leu His Asp Leu Ile His Thr Asp 230 235 240 Arg Ser Leu Thr Leu Val Phe Glu Tyr Leu Asp Ser Asp Leu Lys 245 250 255 Gln Tyr Leu Asp His Cys Gly Asn Leu Met Ser Met His Asn Val 260 265 270 Lys Ile Phe Met Phe Gln Leu Leu Arg Gly Leu Ala Tyr Cys His 275 280 285 His Arg Lys Ile Leu His Arg Asp Leu Lys Pro Gln Asn Leu Leu 290 295 300 Ile Asn Glu Arg Gly Glu Leu Lys Leu Ala Asp Phe Gly Leu Ala 305 310 315 Arg Ala Lys Ser Val Pro Thr Lys Thr Tyr Ser Asn Glu Val Val 320 325 330 Thr Leu Trp Tyr Arg Pro Pro Asp Val Leu Leu Gly Ser Thr Glu 335 340 345 Tyr Ser Thr Pro Ile Asp Met Trp Gly Val Gly Cys Ile His Tyr 350 355 360 Glu Met Ala Thr Gly Arg Pro Leu Phe Pro Gly Ser Thr Val Lys 365 370 375 Glu Glu Leu His Leu Ile Phe Arg Leu Leu Gly Thr Pro Thr Glu 380 385 390 Glu Thr Trp Pro Gly Val Thr Ala Phe Ser Glu Phe Arg Thr Tyr 395 400 405 Ser Phe Pro Cys Tyr Leu Pro Gln Pro Leu Ile Asn His Ala Pro 410 415 420 Arg Leu Asp Thr Asp Gly Ile His Leu Leu Ser Ser Leu Leu Leu 425 430 435 Tyr Glu Ser Lys Ser Arg Met Ser Ala Glu Ala Ala Leu Ser His 440 445 450 Ser Tyr Phe Arg Ser Leu Gly Glu Arg Val His Gln Leu Glu Asp 455 460 465 Thr Ala Ser Ile Phe Ser Leu Lys Glu Ile Gln Leu Gln Lys Asp 470 475 480 Pro Gly Tyr Arg Gly Leu Ala Phe Gln Gln Pro Gly Arg Gly Lys 485 490 495 Asn Arg Arg Gln Ser Ile Phe 500 14 791 PRT Homo sapiens misc_feature Incyte ID No 2192119CD1 14 Met Trp Phe Phe Ala Arg Asp Pro Val Arg Asp Phe Pro Phe Glu 1 5 10 15 Leu Ile Pro Glu Pro Pro Glu Gly Gly Leu Pro Gly Pro Trp Ala 20 25 30 Leu His Arg Gly Arg Lys Lys Ala Thr Gly Ser Pro Val Ser Ile 35 40 45 Phe Val Tyr Asp Val Lys Pro Gly Ala Glu Glu Gln Thr Gln Val 50 55 60 Ala Lys Ala Ala Phe Lys Arg Phe Lys Thr Leu Arg His Pro Asn 65 70 75 Ile Leu Ala Tyr Ile Asp Gly Leu Glu Thr Glu Lys Cys Leu His 80 85 90 Val Val Thr Glu Ala Val Thr Pro Leu Gly Ile Tyr Leu Lys Ala 95 100 105 Arg Val Glu Ala Gly Gly Leu Lys Glu Leu Glu Ile Ser Trp Gly 110 115 120 Leu His Gln Ile Val Lys Ala Leu Ser Phe Leu Val Asn Asp Cys 125 130 135 Ser Leu Ile His Asn Asn Val Cys Met Ala Ala Val Phe Val Asp 140 145 150 Arg Ala Gly Glu Trp Lys Leu Gly Gly Leu Asp Tyr Met Tyr Ser 155 160 165 Ala Gln Gly Asn Gly Gly Gly Pro Pro Arg Lys Gly Ile Pro Glu 170 175 180 Leu Glu Gln Tyr Asp Pro Pro Glu Leu Ala Asp Ser Ser Gly Arg 185 190 195 Val Val Arg Glu Lys Trp Ser Ala Asp Met Trp Arg Leu Gly Cys 200 205 210 Leu Ile Trp Glu Val Phe Asn Gly Pro Leu Pro Arg Ala Ala Ala 215 220 225 Leu Arg Asn Pro Gly Lys Ile Pro Lys Thr Leu Val Pro His Tyr 230 235 240 Cys Glu Leu Val Gly Ala Asn Pro Lys Val Arg Pro Asn Pro Ala 245 250 255 Arg Phe Leu Gln Asn Cys Arg Ala Pro Gly Gly Phe Met Ser Asn 260 265 270 Arg Phe Val Glu Thr Asn Leu Phe Leu Glu Glu Ile Gln Ile Lys 275 280 285 Glu Pro Ala Glu Lys Gln Lys Phe Phe Gln Glu Leu Ser Lys Ser 290 295 300 Leu Asp Ala Phe Pro Glu Asp Phe Cys Arg His Lys Val Leu Pro 305 310 315 Gln Leu Leu Thr Ala Phe Glu Phe Gly Asn Ala Gly Ala Val Val 320 325 330 Leu Thr Pro Leu Phe Lys Val Gly Lys Phe Leu Ser Ala Glu Glu 335 340 345 Tyr Gln Gln Lys Ile Ile Pro Val Val Val Lys Met Phe Ser Ser 350 355 360 Thr Asp Arg Ala Met Arg Ile Arg Leu Leu Gln Gln Met Glu Gln 365 370 375 Phe Ile Gln Tyr Leu Asp Glu Pro Thr Val Asn Thr Gln Ile Phe 380 385 390 Pro His Val Val His Gly Phe Leu Asp Thr Asn Pro Ala Ile Arg 395 400 405 Glu Gln Thr Val Lys Ser Met Leu Leu Leu Ala Pro Lys Leu Asn 410 415 420 Glu Ala Asn Leu Asn Val Glu Leu Met Lys His Phe Ala Arg Leu 425 430 435 Gln Ala Lys Asp Glu Gln Gly Pro Ile Arg Cys Asn Thr Thr Val 440 445 450 Cys Leu Gly Lys Ile Gly Ser Tyr Leu Ser Ala Ser Thr Arg His 455 460 465 Arg Val Leu Thr Ser Ala Phe Ser Arg Ala Thr Arg Asp Pro Phe 470 475 480 Ala Pro Ser Arg Val Ala Gly Val Leu Gly Phe Ala Ala Thr His 485 490 495 Asn Leu Tyr Ser Met Asn Asp Cys Ala Gln Lys Ile Leu Pro Val 500 505 510 Leu Cys Gly Leu Thr Val Asp Pro Glu Lys Ser Val Arg Asp Gln 515 520 525 Ala Phe Lys Ala Ile Arg Ser Phe Leu Ser Lys Leu Glu Ser Val 530 535 540 Ser Glu Asp Pro Thr Gln Leu Glu Glu Val Glu Lys Asp Val His 545 550 555 Ala Ala Ser Ser Pro Gly Met Gly Gly Ala Ala Ala Ser Trp Ala 560 565 570 Gly Trp Ala Val Thr Gly Val Ser Ser Leu Thr Ser Lys Leu Ile 575 580 585 Arg Ser His Pro Thr Thr Ala Pro Thr Glu Thr Asn Ile Pro Gln 590 595 600 Arg Pro Thr Pro Glu Gly His Trp Glu Thr Gln Glu Glu Asp Lys 605 610 615 Asp Thr Ala Glu Asp Ser Ser Thr Ala Asp Arg Trp Asp Asp Glu 620 625 630 Asp Trp Gly Ser Leu Glu Gln Glu Ala Glu Ser Val Leu Ala Gln 635 640 645 Gln Asp Asp Trp Ser Thr Gly Gly Gln Val Ser Arg Ala Ser Gln 650 655 660 Val Ser Asn Ser Asp His Lys Ser Ser Lys Ser Pro Glu Ser Asp 665 670 675 Trp Ser Ser Trp Glu Ala Glu Gly Ser Trp Glu Gln Gly Trp Gln 680 685 690 Glu Pro Ser Ser Gln Glu Pro Pro Pro Asp Gly Thr Arg Leu Ala 695 700 705 Ser Glu Tyr Asn Trp Gly Gly Pro Glu Ser Ser Asp Lys Gly Asp 710 715 720 Pro Phe Ala Thr Leu Ser Ala Arg Pro Ser Thr Gln Pro Arg Pro 725 730 735 Asp Ser Trp Gly Glu Asp Asn Trp Glu Gly Leu Glu Thr Asp Ser 740 745 750 Arg Gln Val Lys Ala Glu Leu Ala Arg Lys Lys Arg Glu Glu Arg 755 760 765 Arg Arg Glu Met Glu Ala Lys Arg Ala Glu Arg Lys Val Ala Lys 770 775 780 Gly Pro Met Lys Leu Gly Ala Arg Lys Leu Asp 785 790 15 1651 PRT Homo sapiens misc_feature Incyte ID No 7474496CD1 15 Met Ala Gly Gly Arg Gly Ala Pro Gly Arg Gly Arg Asp Glu Pro 1 5 10 15 Pro Glu Ser Tyr Pro Gln Arg Gln Asp His Glu Leu Gln Ala Leu 20 25 30 Glu Ala Ile Tyr Gly Ala Asp Phe Gln Asp Leu Arg Pro Asp Ala 35 40 45 Cys Gly Pro Val Lys Glu Pro Pro Glu Ile Asn Leu Val Leu Tyr 50 55 60 Pro Gln Gly Leu Thr Gly Glu Glu Val Tyr Val Lys Val Asp Leu 65 70 75 Arg Val Lys Cys Pro Pro Thr Tyr Pro Asp Val Val Pro Glu Ile 80 85 90 Glu Leu Lys Asn Ala Lys Gly Leu Ser Asn Glu Ser Val Asn Leu 95 100 105 Leu Lys Ser Arg Leu Glu Glu Leu Ala Lys Lys His Cys Gly Glu 110 115 120 Val Met Ile Phe Glu Leu Ala Tyr His Val Gln Ser Phe Leu Ser 125 130 135 Glu His Asn Lys Pro Pro Pro Lys Ser Phe His Glu Glu Met Leu 140 145 150 Glu Arg Arg Ala Gln Glu Glu Gln Gln Arg Leu Leu Glu Ala Gln 155 160 165 Ala Glu Arg Arg Arg Glu Gln Ala Gln Gln Arg Glu Ile Leu His 170 175 180 Glu Ile Gln Arg Arg Lys Glu Glu Ile Lys Glu Glu Lys Lys Arg 185 190 195 Lys Glu Met Ala Lys Gln Glu Arg Leu Glu Ile Ala Ser Leu Ser 200 205 210 Asn Gln Asp His Thr Ser Lys Lys Asp Pro Gly Gly His Arg Thr 215 220 225 Ala Ala Ile Leu His Gly Gly Ser Pro Asp Phe Val Gly Asn Gly 230 235 240 Lys His Arg Ala Asn Ser Ser Gly Arg Ser Arg Arg Glu Arg Gln 245 250 255 Tyr Ser Val Cys Asn Ser Glu Asp Ser Pro Gly Ser Cys Glu Ile 260 265 270 Leu Tyr Phe Asn Met Gly Ser Pro Asp Gln Leu Met Val His Lys 275 280 285 Gly Lys Cys Ile Gly Ser Asp Glu Gln Leu Gly Lys Leu Val Tyr 290 295 300 Asn Ala Leu Glu Thr Ala Thr Gly Gly Phe Val Leu Leu Tyr Glu 305 310 315 Trp Val Leu Gln Trp Gln Lys Lys Met Gly Pro Phe Leu Thr Ser 320 325 330 Gln Glu Lys Glu Lys Ile Asp Lys Cys Lys Lys Gln Ile Gln Gly 335 340 345 Thr Glu Thr Glu Phe Asn Ser Leu Val Lys Leu Ser His Pro Asn 350 355 360 Val Val Arg Tyr Leu Ala Met Asn Leu Lys Glu Gln Asp Asp Ser 365 370 375 Ile Val Val Asp Ile Leu Val Glu His Ile Ser Gly Val Ser Leu 380 385 390 Ala Ala His Leu Ser His Ser Gly Pro Ile Pro Val His Gln Leu 395 400 405 Arg Arg Tyr Thr Ala Gln Leu Leu Ser Gly Leu Asp Tyr Leu His 410 415 420 Ser Asn Ser Val Val His Lys Val Leu Ser Ala Ser Asn Val Leu 425 430 435 Val Asp Ala Glu Gly Thr Val Lys Ile Thr Asp Tyr Ser Ile Ser 440 445 450 Lys Arg Leu Ala Asp Ile Cys Lys Glu Asp Val Phe Glu Gln Thr 455 460 465 Arg Val Arg Phe Ser Asp Asn Ala Leu Pro Tyr Lys Thr Gly Lys 470 475 480 Lys Gly Asp Val Trp Arg Leu Gly Leu Leu Leu Leu Ser Leu Ser 485 490 495 Gln Gly Gln Glu Cys Gly Glu Tyr Pro Val Thr Ile Pro Ser Asp 500 505 510 Leu Pro Ala Asp Phe Gln Asp Phe Leu Lys Cys Val Cys Leu Asp 515

520 525 Asp Lys Glu Arg Trp Ser Pro Gln Gln Leu Leu Lys His Ser Phe 530 535 540 Ile Asn Pro Gln Pro Lys Met Pro Leu Val Glu Gln Ser Pro Glu 545 550 555 Asp Ser Glu Gly Gln Asp Tyr Val Glu Thr Val Ile Pro Ser Asn 560 565 570 Arg Leu Pro Ser Ala Ala Phe Phe Ser Glu Thr Gln Arg Gln Phe 575 580 585 Ser Arg Tyr Phe Ile Glu Phe Glu Glu Leu Gln Leu Leu Gly Lys 590 595 600 Gly Ala Phe Gly Ala Val Ile Lys Val Gln Asn Lys Leu Asp Gly 605 610 615 Cys Cys Tyr Ala Val Lys Arg Ile Pro Ile Asn Pro Ala Ser Arg 620 625 630 Gln Phe Arg Arg Ile Lys Gly Glu Val Thr Leu Leu Ser Arg Leu 635 640 645 His His Glu Asn Ile Val Arg Tyr Tyr Asn Ala Trp Ile Glu Arg 650 655 660 His Glu Arg Pro Ala Gly Pro Gly Thr Pro Pro Pro Asp Ser Gly 665 670 675 Pro Leu Ala Lys Asp Asp Arg Ala Ala Arg Gly Gln Pro Ala Ser 680 685 690 Asp Thr Asp Gly Leu Asp Ser Val Glu Ala Ala Ala Pro Pro Pro 695 700 705 Ile Leu Ser Ser Ser Val Glu Trp Ser Thr Ser Gly Glu Arg Ser 710 715 720 Ala Ser Ala Arg Phe Pro Ala Thr Gly Pro Gly Ser Ser Asp Asp 725 730 735 Glu Asp Asp Asp Glu Asp Glu His Gly Gly Val Phe Ser Gln Ser 740 745 750 Phe Leu Pro Ala Ser Asp Ser Glu Ser Asp Ile Ile Phe Asp Asn 755 760 765 Glu Asp Glu Asn Ser Lys Ser Gln Asn Gln Asp Glu Asp Cys Asn 770 775 780 Glu Lys Asn Gly Cys His Glu Ser Glu Pro Ser Val Thr Thr Glu 785 790 795 Ala Val His Tyr Leu Tyr Ile Gln Met Glu Tyr Cys Glu Lys Ser 800 805 810 Thr Leu Arg Asp Thr Ile Asp Gln Gly Leu Tyr Arg Asp Thr Val 815 820 825 Arg Leu Trp Arg Leu Phe Arg Glu Ile Leu Asp Gly Leu Ala Tyr 830 835 840 Ile His Glu Lys Gly Met Ile His Arg Asp Leu Lys Pro Val Asn 845 850 855 Ile Phe Leu Asp Ser Asp Asp His Val Lys Ile Gly Asp Phe Gly 860 865 870 Leu Ala Thr Asp His Leu Ala Phe Ser Ala Asp Ser Lys Gln Asp 875 880 885 Asp Gln Thr Gly Asp Leu Ile Lys Ser Asp Pro Ser Gly His Leu 890 895 900 Thr Gly Met Val Gly Thr Ala Leu Tyr Val Ser Pro Glu Val Gln 905 910 915 Gly Ser Thr Lys Ser Ala Tyr Asn Gln Lys Val Asp Leu Phe Ser 920 925 930 Leu Gly Ile Ile Phe Phe Glu Met Ser Tyr His Pro Met Val Thr 935 940 945 Ala Ser Glu Arg Ile Phe Val Leu Asn Gln Leu Arg Asp Pro Thr 950 955 960 Ser Pro Lys Phe Pro Glu Asp Phe Asp Asp Gly Glu His Ala Lys 965 970 975 Gln Lys Ser Val Ile Ser Trp Leu Leu Asn His Asp Pro Ala Lys 980 985 990 Arg Pro Thr Ala Thr Glu Leu Leu Lys Ser Glu Leu Leu Pro Pro 995 1000 1005 Pro Gln Met Glu Glu Ser Glu Leu His Glu Val Leu His His Thr 1010 1015 1020 Leu Thr Asn Val Asp Gly Lys Ala Tyr Arg Thr Met Met Ala Gln 1025 1030 1035 Ile Phe Ser Gln Arg Ile Ser Pro Ala Ile Asp Tyr Thr Tyr Asp 1040 1045 1050 Ser Asp Ile Leu Lys Gly Asn Phe Ser Ile Arg Thr Ala Lys Met 1055 1060 1065 Gln Gln His Val Cys Glu Thr Ile Ile Arg Ile Phe Lys Arg His 1070 1075 1080 Gly Ala Val Gln Leu Cys Thr Pro Leu Leu Leu Pro Arg Asn Arg 1085 1090 1095 Gln Ile Tyr Glu His Asn Glu Ala Ala Leu Phe Met Asp His Ser 1100 1105 1110 Gly Met Leu Val Met Leu Pro Phe Asp Leu Arg Ile Pro Phe Ala 1115 1120 1125 Arg Tyr Val Ala Arg Asn Asn Ile Leu Asn Leu Lys Arg Tyr Cys 1130 1135 1140 Ile Glu Arg Val Phe Arg Pro Arg Lys Leu Asp Arg Phe His Pro 1145 1150 1155 Lys Glu Leu Leu Glu Cys Ala Phe Asp Ile Val Thr Ser Thr Thr 1160 1165 1170 Asn Ser Phe Leu Pro Thr Ala Glu Ile Ile Tyr Thr Ile Tyr Glu 1175 1180 1185 Ile Ile Gln Glu Phe Pro Ala Leu Gln Glu Arg Asn Tyr Ser Ile 1190 1195 1200 Tyr Leu Asn His Thr Met Leu Leu Lys Ala Ile Leu Leu His Cys 1205 1210 1215 Gly Ile Pro Glu Asp Lys Leu Ser Gln Val Tyr Ile Ile Leu Tyr 1220 1225 1230 Asp Ala Val Thr Glu Lys Leu Thr Arg Arg Glu Val Glu Ala Lys 1235 1240 1245 Phe Cys Asn Leu Ser Leu Ser Ser Asn Ser Leu Cys Arg Leu Tyr 1250 1255 1260 Lys Phe Ile Glu Gln Lys Gly Asp Leu Gln Asp Leu Met Pro Thr 1265 1270 1275 Ile Asn Ser Leu Ile Lys Gln Lys Thr Gly Ile Ala Gln Leu Val 1280 1285 1290 Lys Tyr Gly Leu Lys Asp Leu Glu Glu Val Val Gly Leu Leu Lys 1295 1300 1305 Lys Leu Gly Ile Lys Leu Gln Val Leu Ile Asn Leu Gly Leu Val 1310 1315 1320 Tyr Lys Val Gln Gln His Asn Gly Ile Ile Phe Gln Phe Val Ala 1325 1330 1335 Phe Ile Lys Arg Arg Gln Arg Ala Val Pro Glu Ile Leu Ala Ala 1340 1345 1350 Gly Gly Arg Tyr Asp Leu Leu Ile Pro Gln Phe Arg Gly Pro Gln 1355 1360 1365 Ala Leu Gly Pro Val Pro Thr Ala Ile Gly Val Ser Ile Ala Ile 1370 1375 1380 Asp Lys Ile Ser Ala Ala Val Leu Asn Met Glu Glu Ser Val Thr 1385 1390 1395 Ile Ser Ser Cys Asp Leu Leu Val Val Ser Val Gly Gln Met Ser 1400 1405 1410 Met Ser Arg Ala Ile Asn Leu Thr Gln Lys Leu Trp Thr Ala Gly 1415 1420 1425 Ile Thr Ala Glu Ile Met Tyr Asp Trp Ser Gln Ser Gln Glu Glu 1430 1435 1440 Leu Gln Glu Tyr Cys Arg His His Glu Ile Thr Tyr Val Ala Leu 1445 1450 1455 Val Ser Asp Lys Glu Gly Ser His Val Lys Val Lys Ser Phe Glu 1460 1465 1470 Lys Glu Arg Gln Thr Glu Lys Arg Val Leu Glu Thr Glu Leu Val 1475 1480 1485 Asp His Val Leu Gln Lys Leu Arg Thr Lys Val Thr Asp Glu Arg 1490 1495 1500 Asn Gly Arg Glu Ala Ser Asp Asn Leu Ala Val Gln Asn Leu Lys 1505 1510 1515 Gly Ser Phe Ser Asn Ala Ser Gly Leu Phe Glu Ile His Gly Ala 1520 1525 1530 Thr Val Val Pro Ile Val Ser Val Leu Ala Pro Glu Lys Leu Ser 1535 1540 1545 Ala Ser Thr Arg Arg Arg Tyr Glu Thr Gln Val Gln Thr Arg Leu 1550 1555 1560 Gln Thr Ser Leu Ala Asn Leu His Gln Lys Ser Ser Glu Ile Glu 1565 1570 1575 Ile Leu Ala Val Asp Leu Pro Lys Glu Thr Ile Leu Gln Phe Leu 1580 1585 1590 Ser Leu Glu Trp Asp Ala Asp Glu Gln Ala Phe Asn Thr Thr Val 1595 1600 1605 Lys Gln Leu Leu Ser Arg Leu Pro Lys Gln Arg Tyr Leu Lys Leu 1610 1615 1620 Val Cys Asp Glu Ile Tyr Asn Ile Lys Val Glu Lys Lys Val Ser 1625 1630 1635 Val Leu Phe Leu Tyr Ser Tyr Arg Asp Asp Tyr Tyr Arg Ile Leu 1640 1645 1650 Phe 16 752 PRT Homo sapiens misc_feature Incyte ID No 1834248CD1 16 Met Ser Ser Arg Thr Val Leu Ala Pro Gly Asn Asp Arg Asn Ser 1 5 10 15 Asp Thr His Gly Thr Leu Gly Ser Gly Arg Ser Ser Asp Lys Gly 20 25 30 Pro Ser Trp Ser Ser Arg Ser Leu Gly Ala Arg Cys Arg Asn Ser 35 40 45 Ile Ala Ser Cys Pro Glu Glu Gln Pro His Val Gly Asn Tyr Arg 50 55 60 Leu Leu Arg Thr Ile Gly Lys Gly Asn Phe Ala Lys Val Lys Leu 65 70 75 Ala Arg His Ile Leu Thr Gly Arg Glu Val Ala Ile Lys Ile Ile 80 85 90 Asp Lys Thr Gln Leu Asn Pro Ser Ser Leu Gln Lys Leu Phe Arg 95 100 105 Glu Val Arg Ile Met Lys Gly Leu Asn His Pro Asn Ile Val Lys 110 115 120 Leu Phe Glu Val Ile Glu Thr Glu Lys Thr Leu Tyr Leu Val Met 125 130 135 Glu Tyr Ala Ser Ala Gly Glu Val Phe Asp Tyr Leu Val Ser His 140 145 150 Gly Arg Met Lys Glu Lys Glu Ala Arg Ala Lys Phe Arg Gln Ile 155 160 165 Val Ser Ala Val His Tyr Cys His Gln Lys Asn Ile Val His Arg 170 175 180 Asp Leu Lys Ala Glu Asn Leu Leu Leu Asp Ala Glu Ala Asn Ile 185 190 195 Lys Ile Ala Asp Phe Gly Phe Ser Asn Glu Phe Thr Leu Gly Ser 200 205 210 Lys Leu Asp Thr Phe Cys Gly Ser Pro Pro Tyr Ala Ala Pro Glu 215 220 225 Leu Phe Gln Gly Lys Lys Tyr Asp Gly Pro Glu Val Asp Ile Trp 230 235 240 Ser Leu Gly Val Ile Leu Tyr Thr Leu Val Ser Gly Ser Leu Pro 245 250 255 Phe Asp Gly His Asn Leu Lys Glu Leu Arg Glu Arg Val Leu Arg 260 265 270 Gly Lys Tyr Arg Val Pro Phe Tyr Met Ser Thr Asp Cys Glu Ser 275 280 285 Ile Leu Arg Arg Phe Leu Val Leu Asn Pro Ala Lys Arg Cys Thr 290 295 300 Leu Glu Gln Ile Met Lys Asp Lys Trp Ile Asn Ile Gly Tyr Glu 305 310 315 Gly Glu Glu Leu Lys Pro Tyr Thr Glu Pro Glu Glu Asp Phe Gly 320 325 330 Asp Thr Lys Arg Ile Glu Val Met Val Gly Met Gly Tyr Thr Arg 335 340 345 Glu Glu Ile Lys Glu Ser Leu Thr Ser Gln Lys Tyr Asn Glu Val 350 355 360 Thr Ala Thr Tyr Leu Leu Leu Gly Arg Lys Thr Glu Glu Gly Gly 365 370 375 Asp Arg Gly Ala Pro Gly Leu Ala Leu Ala Arg Val Arg Ala Pro 380 385 390 Ser Asp Thr Thr Asn Gly Thr Ser Ser Ser Lys Gly Thr Ser His 395 400 405 Ser Lys Gly Gln Arg Ser Ser Ser Ser Thr Tyr His Arg Gln Arg 410 415 420 Arg His Ser Asp Phe Cys Gly Pro Ser Pro Ala Pro Leu His Pro 425 430 435 Lys Arg Ser Pro Thr Ser Thr Gly Glu Ala Glu Leu Lys Glu Glu 440 445 450 Arg Leu Pro Gly Arg Lys Ala Ser Cys Ser Thr Ala Gly Ser Gly 455 460 465 Ser Arg Gly Leu Pro Pro Ser Ser Pro Met Val Ser Ser Ala His 470 475 480 Asn Pro Asn Lys Ala Glu Ile Pro Glu Arg Arg Lys Asp Ser Thr 485 490 495 Ser Thr Pro Asn Asn Leu Pro Pro Ser Met Met Thr Arg Arg Asn 500 505 510 Thr Tyr Val Cys Thr Glu Arg Pro Gly Ala Glu Arg Pro Ser Leu 515 520 525 Leu Pro Asn Gly Lys Glu Asn Ser Ser Gly Thr Pro Arg Val Pro 530 535 540 Pro Ala Ser Pro Ser Ser His Ser Leu Ala Pro Pro Ser Gly Glu 545 550 555 Arg Ser Arg Leu Ala Arg Gly Ser Thr Ile Arg Ser Thr Phe His 560 565 570 Gly Gly Gln Val Arg Asp Arg Arg Ala Gly Gly Gly Gly Gly Gly 575 580 585 Gly Val Gln Asn Gly Pro Pro Ala Ser Pro Thr Leu Ala His Glu 590 595 600 Ala Ala Pro Leu Pro Ala Gly Arg Pro Arg Pro Thr Thr Asn Leu 605 610 615 Phe Thr Lys Leu Thr Ser Lys Leu Thr Arg Arg Val Ala Asp Glu 620 625 630 Pro Glu Arg Ile Gly Gly Pro Glu Val Thr Ser Cys His Leu Pro 635 640 645 Trp Asp Gln Thr Glu Thr Ala Pro Arg Leu Leu Arg Phe Pro Trp 650 655 660 Ser Val Lys Leu Thr Ser Ser Arg Pro Pro Glu Ala Leu Met Ala 665 670 675 Ala Leu Arg Gln Ala Thr Ala Ala Ala Arg Cys Arg Cys Arg Gln 680 685 690 Pro Gln Pro Phe Leu Leu Ala Cys Leu His Gly Gly Ala Gly Gly 695 700 705 Pro Glu Pro Leu Ser His Phe Glu Val Glu Val Cys Gln Leu Pro 710 715 720 Arg Pro Gly Leu Arg Gly Val Leu Phe Arg Arg Val Ala Gly Thr 725 730 735 Ala Leu Ala Phe Arg Thr Leu Val Thr Arg Ile Ser Asn Asp Leu 740 745 750 Glu Leu 17 501 PRT Homo sapiens misc_feature Incyte ID No 71584520CD1 17 Met Pro Phe Gly Cys Val Thr Leu Gly Asp Lys Lys Asn Tyr Asn 1 5 10 15 Gln Pro Ser Glu Val Thr Asp Arg Tyr Asp Leu Gly Gln Val Ile 20 25 30 Lys Thr Glu Glu Phe Cys Glu Ile Phe Arg Ala Lys Asp Lys Thr 35 40 45 Thr Gly Lys Leu His Thr Cys Lys Lys Phe Gln Lys Arg Asp Gly 50 55 60 Arg Lys Val Arg Lys Ala Ala Lys Asn Glu Ile Gly Ile Leu Lys 65 70 75 Met Val Lys His Pro Asn Ile Leu Gln Leu Val Asp Val Phe Val 80 85 90 Thr Arg Lys Glu Tyr Phe Ile Phe Leu Glu Leu Ala Thr Gly Arg 95 100 105 Glu Val Phe Asp Trp Ile Leu Asp Gln Gly Tyr Tyr Ser Glu Arg 110 115 120 Asp Thr Ser Asn Val Val Arg Gln Val Leu Glu Ala Val Ala Tyr 125 130 135 Leu His Ser Leu Lys Ile Val His Arg Asn Leu Lys Leu Glu Asn 140 145 150 Leu Val Tyr Tyr Asn Arg Leu Lys Asn Ser Lys Ile Val Ile Ser 155 160 165 Asp Phe His Leu Ala Lys Leu Glu Asn Gly Leu Ile Lys Glu Pro 170 175 180 Cys Gly Thr Pro Glu Tyr Leu Ala Pro Glu Val Val Gly Arg Gln 185 190 195 Arg Tyr Gly Arg Pro Val Asp Cys Trp Ala Ile Gly Val Ile Met 200 205 210 Tyr Ile Leu Leu Ser Gly Asn Pro Pro Phe Tyr Glu Glu Val Glu 215 220 225 Glu Asp Asp Tyr Glu Asn His Asp Lys Asn Leu Phe Arg Lys Ile 230 235 240 Leu Ala Gly Asp Tyr Glu Phe Asp Ser Pro Tyr Trp Asp Asp Ile 245 250 255 Ser Gln Ala Ala Lys Asp Leu Val Thr Arg Leu Met Glu Val Glu 260 265 270 Gln Asp Gln Arg Ile Thr Ala Glu Glu Ala Ile Ser His Glu Trp 275 280 285 Ile Ser Gly Asn Ala Ala Ser Asp Lys Asn Ile Lys Asp Gly Val 290 295 300 Cys Ala Gln Ile Glu Lys Asn Phe Ala Arg Ala Lys Trp Lys Lys 305 310 315 Ala Val Arg Val Thr Thr Leu Met Lys Arg Leu Arg Ala Pro Glu 320 325 330 Gln Ser Ser Thr Ala Ala Ala Gln Ser Ala Ser Ala Thr Asp Thr 335 340 345 Ala Thr Pro Gly Ala Ala Gly Gly Ala Thr Ala Ala Ala Ala Ser 350 355 360 Gly Ala Thr Ser Ala Pro Glu Gly Asp Ala Ala Arg Ala Ala Lys 365 370 375 Ser Asp Asn Val Ala Pro Ala Asp Arg Ser Ala Thr Pro Ala Thr 380 385 390 Asp Gly

Ser Ala Thr Pro Ala Thr Asp Gly Ser Val Thr Pro Ala 395 400 405 Thr Asp Gly Ser Ile Thr Pro Ala Thr Asp Gly Ser Val Thr Pro 410 415 420 Ala Thr Asp Arg Ser Ala Thr Pro Ala Thr Asp Gly Arg Ala Thr 425 430 435 Pro Ala Thr Glu Glu Ser Thr Val Pro Thr Thr Gln Ser Ser Ala 440 445 450 Met Leu Ala Thr Lys Ala Ala Ala Thr Pro Glu Pro Ala Met Ala 455 460 465 Gln Pro Asp Ser Thr Ala Pro Glu Gly Ala Thr Gly Gln Ala Pro 470 475 480 Pro Ser Ser Lys Gly Glu Glu Ala Ala Gly Tyr Ala Gln Glu Ser 485 490 495 Gln Arg Glu Glu Ala Ser 500 18 346 PRT Homo sapiens misc_feature Incyte ID No 7475538CD1 18 Met Asp Gln Tyr Cys Ile Leu Gly Arg Ile Gly Glu Gly Ala His 1 5 10 15 Gly Ile Val Phe Lys Ala Lys His Val Glu Thr Gly Glu Ile Val 20 25 30 Ala Leu Lys Lys Val Ala Leu Arg Arg Leu Glu Asp Gly Phe Pro 35 40 45 Asn Gln Ala Leu Arg Glu Ile Lys Ala Leu Gln Glu Met Glu Asp 50 55 60 Asn Gln Tyr Val Val Gln Leu Lys Ala Val Phe Pro His Gly Gly 65 70 75 Gly Phe Val Leu Ala Phe Glu Phe Met Leu Ser Asp Leu Ala Glu 80 85 90 Val Val Arg His Ala Gln Arg Pro Leu Ala Gln Ala Gln Val Lys 95 100 105 Ser Tyr Leu Gln Met Leu Leu Lys Gly Val Ala Phe Cys His Ala 110 115 120 Asn Asn Ile Val His Arg Asp Leu Lys Pro Ala Asn Leu Leu Ile 125 130 135 Ser Ala Ser Gly Gln Leu Lys Ile Ala Asp Phe Gly Leu Ala Arg 140 145 150 Val Phe Ser Pro Asp Gly Ser Arg Leu Tyr Thr His Gln Val Ala 155 160 165 Thr Arg Trp Tyr Arg Ala Pro Glu Leu Leu Tyr Gly Ala Arg Gln 170 175 180 Tyr Asp Gln Gly Val Asp Leu Trp Ser Val Gly Cys Ile Met Gly 185 190 195 Glu Leu Leu Asn Gly Ser Pro Leu Phe Pro Gly Lys Asn Asp Ile 200 205 210 Glu Gln Leu Cys Tyr Val Leu Arg Ile Leu Gly Thr Pro Asn Pro 215 220 225 Gln Val Trp Pro Glu Leu Thr Glu Leu Pro Asp Tyr Asn Lys Ile 230 235 240 Ser Phe Lys Glu Gln Val Pro Met Pro Leu Glu Glu Val Leu Pro 245 250 255 Asp Val Ser Pro Gln Ala Leu Asp Leu Leu Gly Gln Phe Leu Leu 260 265 270 Tyr Pro Pro His Gln Arg Ile Ala Ala Ser Lys Ala Leu Leu His 275 280 285 Gln Tyr Phe Phe Thr Ala Pro Leu Pro Ala His Pro Ser Glu Leu 290 295 300 Pro Ile Pro Gln Arg Leu Gly Gly Pro Ala Pro Lys Ala His Pro 305 310 315 Gly Pro Pro His Ile His Asp Phe His Val Asp Arg Pro Leu Glu 320 325 330 Glu Ser Leu Leu Asn Pro Glu Leu Ile Arg Pro Phe Ile Leu Glu 335 340 345 Gly 19 4224 DNA Homo sapiens misc_feature Incyte ID No 2890544CB1 19 ccggcggtag ccccggacgg cagcaggagg ccgaggcggg agcgcgcggg gctgaggcgg 60 cggcggcggc ggcgggaagg ggctgggtgg tggagcggga gggaggctga ggaggctccc 120 cctgcgggac gggcgcgggg acggctccgg ggggcggggg ccggggcccg ggccggctcg 180 cgcggggggt atgatgaccc ggctgcgggg ccccagatct cgtctcctcc gccgcctcct 240 cacggcagcc ccagctcgcg gctgagaacc agacacaccg gcggtatggc atcacagctg 300 caagtgtttt cgcccccatc agtgtcgtcg agtgccttct gcagtgcgaa gaaactgaaa 360 atagagccct ctggctggga tgtttcagga cagagtagca acgacaaata ttatacccac 420 agcaaaaccc tcccagccac acaagggcaa gccaactcct ctcaccaggt agcaaatttc 480 aacatccctg cttacgacca gggcctcctc ctcccagctc ctgcagtgga gcatattgtt 540 gtaacagccg ctgatagctc gggcagtgct gctacatcaa ccttccaaag cagccagacc 600 ctgactcaca gaagcaacgt ttctttgctt gagccatatc aaaaatgtgg attgaaacga 660 aaaagtgagg aagttgacag caacggtagt gtgcagatca tagaagaaca tccccctctc 720 atgctgcaaa acaggactgt ggtgggtgct gctgccacaa ccaccactgt gaccacaaag 780 agtagcagtt ccagcggaga aggggattac cagctggtcc agcatgagat cctttgctct 840 atgaccaata gctatgaagt cttggagttc ctaggccggg ggacatttgg acaggtggct 900 aagtgctgga agaggagcac caaggaaatt gtggctatta aaatcttgaa gaaccacccc 960 tcctatgcca gacaaggaca gattgaagtg agcatccttt cccgcctaag cagtgaaaat 1020 gctgatgagt ataatttggt ccgttcatac gagtgctttc agcataagaa tcacacctgc 1080 cttgtttttg aaatgttgga gcagaactta tatgattttc taaagcaaaa caaatttagc 1140 ccactgccac tcaagtacat cagaccaatc ttgcagcagg tggccacagc cttgatgaag 1200 ctcaagagtc ttggtctgat ccacgctgac cttaagcctg aaaacatcat gctggttgat 1260 ccagttcgcc agccctaccg agtgaaggtc attgactttg gttctgctag tcacgtttcc 1320 aaagctgtgt gctcaaccta cttacagtca cgttactaca gagctcctga aattattctt 1380 gggttaccat tttgtgaagc tattgatatg tggtcactgg gctgtgtgat agctgagctg 1440 ttcctgggat ggcctcttta tcctggtgct tcagaatatg atcagattcg ttatatttca 1500 caaacacaag gcttgccagc tgaatatctt ctcagtgccg gaacaaaaac aaccaggttt 1560 ttcaacagag atcctaattt ggggtaccca ctgtggaggc ttaagacacc tgaagaacat 1620 gaactggaga ctggaataaa atcaaaagaa gctcggaagt acatttttaa ttgcttagat 1680 gacatggctc aggtgaatat gtctacagac ctggagggaa cagacatgtt ggcagagaag 1740 gcagaccgaa gagaatacat tgatctgtta aagaaaatgc tcacaattga tgcagataag 1800 agaattaccc ctctaaaaac tcttaaccat cagtttgtga caatgactca ccttttggat 1860 tttccacata gcaatcatgt taagtcttgt tttcagaaca tggagatctg caagcggagg 1920 gttcacatgt atgatacagt gagtcagatc aagagtccct tcactacaca tgttgcccca 1980 aatacaagca caaatctaac catgagcttc agcaatcagc tcaatacagt gcacaatcag 2040 gccagtgttc tagcttccag ttctactgca gcagctgcta ctctttctct ggctaattca 2100 gatgtctcac tactaaacta ccagtcagct ttgtacccat catctgctgc accagttcct 2160 ggagttgccc agcagggtgt ttccttgcag cctggaacca cccagatttg cactcagaca 2220 gatccattcc aacagacatt tatagtatgt ccacctgcgt ttcaaactgg actacaagca 2280 acaacaaagc attctggatt ccctgtgagg atggataatg ctgtaccgat tgtaccccag 2340 gcaccagctg ctcagccact acagattcag tcaggagttc tcacgcaggg aagctgtaca 2400 ccactaatgg tagcaactct ccaccctcaa gtagccacca tcacaccgca gtatgcggtg 2460 ccctttactc tgagctgcgc agccggccgg ccggcgctgg ttgaacagac tgccgctgta 2520 ctgcaggcgt ggcctggagg gactcagcaa attctcctgc cttcaacttg gcaacagttg 2580 cctggggtag ctctacacaa ctctgtccag cccacagcaa tgattccaga ggccatgggg 2640 agtggacagc agctagctga ctggaggaat gcccactctc atggcaacca gtacagcact 2700 atcatgcagc agccatcctt gctgactaac catgtgacat tggccactgc tcagcctctg 2760 aatgttggtg ttgcccatgt tgtcagacaa caacaatcca gttccctccc ttcgaagaag 2820 aataagcagt cagctccagt ctcttccaag tcctctctag atgttctgcc ttcccaagtc 2880 tattctctgg ttgggagcag tcccctccgc accacatctt cttataattc cttggtccct 2940 gtccaagatc agcatcagcc catcatcatt ccagatactc ccagccctcc tgtgagtgtc 3000 atcactatcc gaagtgacac tgatgaggaa gaggacaaca aatacaagcc cagtagctct 3060 ggactgaagc caaggtctaa tgtcatcagt tatgtcactg tcaatgattc tccagactct 3120 gactcttctt tgagcagccc ttattccact gataccctga gtgctctccg aggcaatagt 3180 ggatccgttt tggaggggcc tggcagagtt gtggcagatg gcactggcac ccgcactatc 3240 attgtgcctc cactgaaaac tcagcttggt gactgcactg tagcaaccca ggcctcaggt 3300 ctcctgagca ataagactaa gccagtcgct tcagtgagtg ggcagtcatc tggatgctgt 3360 atcaccccca cagggtatcg agctcaacgc ggggggacca gtgcagcaca accactcaat 3420 cttagccaga accagcagtc atcggcggct ccaacctcac aggagagaag cagcaaccca 3480 gccccccgca ggcagcaggc gtttgtggcc cctctctccc aagcccccta caccttccag 3540 catggcagcc cgctacactc gacagggcac ccacaccttg ccccggcccc tgctcacctg 3600 ccaagccagg ctcatctgta tacgtatgct gccccgactt ctgctgctgc actgggctca 3660 accagctcca ttgctcatct tttctcccca cagggttcct caaggcatgc tgcagcctat 3720 accactcacc ctagcacttt ggtgcaccag gtccctgtca gtgttgggcc cagcctcctc 3780 acttctgcca gcgtggcccc tgctcagtac caacaccagt ttgccaccca atcctacatt 3840 gggtcttccc gaggctcaac aatttacact ggatacccgc tgagtcctac caagatcagc 3900 cagtattcct acttatagtt ggtgagcatg agggaggagg aatcatggct accttctcct 3960 ggccctgcgt tcttaatatt gggctatgga gagatcctcc tttaccctct tgaaatttct 4020 tagccagcaa cttgttctgc aggggcccac tgaagcagaa ggtttttctc tgggggaacc 4080 tgtctcagtg ttgactgcat tgttgtagtc ttctcaaagt tggccctatt tttaaatcca 4140 tatttttgtg acgtcattcg gtactggaga gtcaatgcca tttctgcgtc cagcagaagt 4200 gctcgattcc ctgaatttgc cctg 4224 20 1736 DNA Homo sapiens misc_feature Incyte ID No 7472693CB1 20 gcggcggcgg cgagagcgaa agaggaaact gcagaggagg aagctgcgcc gcagcccgag 60 ccgcccggca tccccgccgc ctctgcgccc gcgccgcgcc cccggcgccc cctccccagc 120 gcgcccccgg ccgctcctcc gcgccgcgct cgtcggccat ggcccgggag aacggcgaga 180 gcagctcctc ctggaaaaag caagctgaag acatcaagaa gatcttcgag ttcaaagaga 240 ccctcggaac cggggccttt tccgaagtgg ttttagctga agagaaggca actggcaagc 300 tctttgctgt gaagtgtatc cctaagaagg cgctgaaggg caaggaaagc agcatagaga 360 atgagatagc cgtcctgaga aagattaagc atgaaaatat tgttgccctg gaagacattt 420 atgaaagccc aaatcacctg tacttggtca tgcagctggt gtccggtgga gagctgtttg 480 accggatagt ggagaagggg ttttatacag agaaggatgc cagcactctg atccgccaag 540 tcttggacgc cgtgtactat ctccacagaa tgggcatcgt ccacagagac ctcaagcccg 600 aaaatctctt gtactacagt caagatgagg agtccaaaat aatgatcagt gactttggat 660 tgtcaaaaat ggagggcaaa ggagatgtga tgtccactgc ctgtggaact ccaggctatg 720 tcgctcctga agtcctcgcc cagaaacctt acagcaaagc cgttgactgc tggtccatcg 780 gagtgattgc ctacatcttg ctctgcggct accctccttt ttatgatgaa aatgactcca 840 agctctttga gcagatcctc aaggcggaat atgagtttga ctctccctac tgggatgaca 900 tctccgactc tgcaaaagac ttcattcgga acctgatgga gaaggacccg aataaaagat 960 acacgtgtga gcaggcagct cggcacccat ggatcgctgg tgacacagcc ctcaacaaaa 1020 acatccacga gtccgtcagc gcccagatcc ggaaaaactt tgccaagagc aaatggagac 1080 aagcatttaa tgccacggcc gtcgtgagac atatgagaaa actacacctc ggcagcagcc 1140 tggacagttc aaatgcaagt gtttcgagca gcctcagttt ggccagccaa aaagactgtg 1200 cgtatgtagc aaaaccagaa tccctcagct gacactgaag acgagcctgg ggtggagagg 1260 agggagccgg catctgccga gcacctcctg tttgccaggc gctttctata cttaatccca 1320 tgtcatgcga ccctaggact ttttttaaca tgtaatcact gggccgggtg cagtggctca 1380 cgcctgtaat cccaacactt tgggaggctg aggcaggagg actgtttgag ttcaggagtt 1440 ttaagaccag cctgaccaac atggtgaaac cccatctcta ctaaaatata aaaattagcc 1500 gggtgtggtg gcgagcacct gtaatgtcag ctacttggga ggctgaggca ggagaatcac 1560 ttgaacccag gaagcggagg ttgcaatgag ctgagatcac accactgcac tccagcctgg 1620 gtgacagatt gagactccct ctcaaaaaaa aaagggaaat cattgaacac tcgtggaacc 1680 ctaggtattg catattccat ttacggtttg ggaatccagg gctcaagtcc tcgcag 1736 21 1824 DNA Homo sapiens misc_feature Incyte ID No 3107952CB1 21 cacggcttcc ggtgtcatgg ctgcttgaag tcccgggagt cggtgaggcg gctgcaggtc 60 cctccctgcg gagccgctgg tccggctggc ggagatgtga ccgcgggccc ggccggcctg 120 cctcaggcgt cgcgtcagct cccgtgtccg tgcccttaac ccacaccgat ggcgggatcc 180 ggctgcgcct ggggcgcgga gccgccgcgt tttctggagg ccttcgggcg gctgtggcag 240 gtacagagcc gtctgggtag cggctcctcc gcctcggtgt atcgggttcg ctgctgcggc 300 aaccctggct cgccccccgg cgccctcaag cagttcttgc cgccaggaac caccggggct 360 gcggcctctg ccgccgagta tggtttccgc aaagagaggg cggcgctgga acagttgcag 420 ggtcacagaa acatcgtgac tttgtatgga gtgtttacaa tccacttttc tccaaatgtg 480 ccatcacgct gtctgttgct tgaactcctg gatgtcagtg tttcggaatt gctcttatat 540 tccagtcacc agggttgttc catgtggatg atacagcatt gtgcccgaga tgttttggag 600 gcccttgctt ttcttcatca tgagggctat gtccatgcgg acctcaaacc acgtaacata 660 ttgtggagtg cagagaatga atgttttaaa ctcattgact ttggacttag cttcaaagaa 720 ggcaatcagg atgtaaagta tattcagaca gacgggtatc gggctccaga agcagaattg 780 caaaattgct tggcccaggc tggcctgcag agtgatacag aatgtacctc agctgttgat 840 ctgtggagcc taggaatcat tttactggaa atgttctcag gaatgaaact gaaacataca 900 gtcagatctc aggaatggaa ggcaaacagt tctgctatta ttgatcacat atttgccagt 960 aaagcagtgg tgaatgccgc aattccagcc tatcacctaa gagaccttat caaaagcatg 1020 cttcatgatg atccaagcag aagaattcct gctgaaatgg cattgtgcag cccattcttt 1080 agcattcctt ttgcccctca tattgaagat ctggtcatgc ttcccactcc agtgctaaga 1140 ctgctgaatg tgctggatga tgattatctt gagaatgaag aggaatatga agatgttgta 1200 gaagatgtaa aagaggagtg tcaaaaatat ggaccagtgg tatctctact tgttccaaag 1260 ggaaatcctg gcagaggaca agtctttgtt gagtatgcaa atgctggtga ttccaaagct 1320 gcgcagaaat tactgactgg aaggatgttt gatgggaagt ttgttgtggc tacattctac 1380 ccgctgagtg cctacaagag gggatatctg tatcaaacct tgctttaatc agtaacctaa 1440 ggactgtttc ctttttctcc tcttccattt cttgggttat tccacatatg aatgcaggac 1500 taccccctta ccattttaag aaggtacttt atacatttat ttaatcctac taatgtgcag 1560 ccattgccca agcagtgact gcgttgcata catttggcac tgagtaggac aagacctctc 1620 agctatacat tgaggggttt tagagcatcc atgtgggcaa cccttttttg tgcgggagag 1680 caggtgttgc tcttcagtat gtagcctaaa aaaatcttaa ttatttcatg gatcatgaag 1740 caaggatgaa taatatcatg tcttggtaaa tactaacaaa tttgttaggt ttggtgacat 1800 catttacagg ttatttcctt atgt 1824 22 2201 DNA Homo sapiens misc_feature Incyte ID No 5544420CB1 22 atgaaccgat acacaaccat gagacagttg ggggacggca cgtatgggag tgtgcttatg 60 ggcaagagta atgaatccgg ggagctggtg gccatcaaaa ggatgaagag aaagttctat 120 tcttgggatg aatgcatgaa cttgagagaa gttaagtctc tgaagaaact taatcatgcc 180 aatgttatta aattgaaaga agttatcaga gaaaatgacc atctttattt tatatttgaa 240 tatatgaaag aaaacctcta tcaattaatg aaagacagga acaagttgtt ccctgaatca 300 gtcatcagaa atattatgta tcaaatattg caagggctgg cttttatcca taaacatggt 360 ttttttcata gggacatgaa accagaaaac ttgctttgta tgggtccaga gcttgtgaaa 420 attgctgatt ttggacttgc aagagaatta aggtcacagc caccatacac tgattatgta 480 tctaccagat ggtatcgtgc ccctgaagtt ttactgagat cttcagttta tagttctccc 540 attgatgtgt gggctgttgg aagtatcatg gctgaactct atatgttaag gccacttttc 600 ccagggacaa gtgaggtcga tgaaatcttt aaaatttgcc aagttttagg gactcccaaa 660 aaaagtgact ggccagaagg ataccagctg gcatcctcta tgaacttccg ttttccccag 720 tgtgttccta taaacttaaa aactcttatt cccaatgcca gtaatgaagc tattcagctc 780 atgaccgaaa tgttgaattg ggatccaaag aaacgaccga cagcaagcca ggcattgaaa 840 cacccatatt ttcaagttgg tcaggtatta ggcccttcgt caaatcatct ggaatcaaaa 900 cagtctttaa ataagcagct gcaaccatta gaatcaaagc catctttagt tgaggtagag 960 cctaagcctc tgccggatat aatcgatcag gttgttggac aaccccagcc aaaaactagc 1020 cagcagccac tgcagcccat tcagccgcca cagaacctga gcgtccagca acctccaaag 1080 caacagagtc aggagaaacc gccacaaacg ctattcccga gcatcgtcaa aaacatgcca 1140 actaagccaa atggcacact gagtcataaa agtggtagga ggcgttgggg tcagactatc 1200 ttcaagtctg gagatagctg ggaagagttg gaggactatg atttcggagc ctcccattcc 1260 aagaagccaa gcatgggtgt ttttaaagaa aaaaggaaaa aagattctcc atttcggctt 1320 ccagagccag taccctcagg ctccaaccac tcgacagggg aaaacaagag cttacctgct 1380 gttacttccc taaaatctga ttccgaattg tcaactgctc caacctctaa acagtactac 1440 ttgaaacaat caagatatct tccaggtgtg aatcccaaga aggtgtcctt gatagccagt 1500 ggaaaggaaa taaaccccca cacttggagc aaccagttat tccccaagtc actgggaccc 1560 gttggggcag aacttgcttt caaaaggagc aatgcagaag aaaaacttgg aagttatgct 1620 acttacaatc agtcaggata tattccttcc tttctcaaaa aagaagtgca gtcagctggc 1680 cagaggatcc acttagcacc tctcaatgca acggcttcag aatatacctg gaacacaaaa 1740 actggtcggg ggcagttttc aggacgtact tataatccta cagcaaaaaa cctaaatatt 1800 gtgaaccgtg cacagcccat tccctcagtg cacgggagga cagactgggt ggccaagtat 1860 ggaggccacc ggtaggagtc tatggtgtga aaccctacag cattgctccg tagagtacgt 1920 gcaagttcct tgaccctggg aaatgtctac aaatgtctat ttctactgag ttctggaaga 1980 aatatgcaaa agtgggtact tggaagggca aaaatcatcc cctattttac ttatttccaa 2040 gaaatgcatt ttcttagcat cattgcccac agtgttgata tatgggtagg atgttacaaa 2100 gtattgaata aactatttgc caaagtatga agtatttgat ctacaattta ataaatagta 2160 aatccaataa gaacccttaa aaaaaaacaa cttccagaaa a 2201 23 2974 DNA Homo sapiens misc_feature Incyte ID No 7472832CB1 23 gcgagatccc gcggatctag aacccagtgt cccccggggc cccccggccg ggtcccgggt 60 gggctccagg cggccggtcc ccggcctccc cccatggcca ccgccccctc ttatcccgcc 120 gggctccctg gctctcccgg gccggggtct cctccgcccc ccggcggcct agagctgcag 180 tcgccgccac cgctactgcc ccagatcccg gccccgggtt ccggggtctc ctttcacatc 240 cagatcgggc tgacccgcga gttcgtgctg ttgcccgccg cctccgagct ggctcatgtg 300 aagcagctgg cctgttccat cgtggaccag aagttccctg agtgtggctt ctacggcctt 360 tacgacaaga tcctgctttt caaacatgac cccacgtcgg ccaacctcct gcagctggtg 420 cgctcgtccg gagacatcca ggagggcgac ctggtggagg tggtgctgtc ggcctcggcc 480 accttcgagg acttccagat ccgcccgcac gccctcacgg tgcactccta tcgggcgcct 540 gccttctgtg atcactgcgg ggagatgctc ttcggcctag tgcgccaggg cctcaagtgc 600 gatggctgcg ggctgaacta ccacaagcgc tgtgccttca gcatccccaa caactgtagt 660 ggggcccgca aacggcgcct gtcatccacg tctctggcca gtggccactc ggtgcgcctc 720 ggcacctccg agtccctgcc ctgcacggct gaagagctga gccgtagcac caccgaactc 780 ctgcctcgcc gtcccccgtc atcctcttcc tcctcttctg cctcatcgta tacgggccgc 840 cccattgagc tggacaagat gctgctctcc aaggtcaagg tgccgcacac cttcctcatc 900 cacagctata cacggcccac cgtttgccag gcttgcaaga aactcctcaa gggcctcttc 960 cggcagggcc tgcaatgcaa agactgcaag tttaactgtc acaaacgctg cgccacccgc 1020 gtccctaatg actgcctggg ggaggccctt atcaatggag atgtgccgat ggaggaggcc 1080 accgatttca gcgaggctga caagagcgcc ctcatggatg agtcagagga ctccggtgtc 1140 atccctggct cccactcaga gaatgcgctc cacgccagtg aggaggagga aggcgaggga 1200 ggcaaggccc agagctccct ggggtacatc cccctaatga gggtggtgca atcggtgcga 1260 cacacgacgc ggaaatccag caccacgctg cgggagggtt gggtggttca ttacagcaac 1320 aaggacacgc tgagaaagcg gcactattgg cgcctggact gcaagtgtat cacgctcttc 1380 cagaacaaca cgaccaacag atactataag gaaattccgc tgtcagaaat cctcacggtg 1440 gagtccgccc agaacttcag ccttgtgccg ccgggcacca acccacactg ctttgagatc 1500 gtcactgcca atgccaccta cttcgtgggc gagatgcctg gcgggactcc gggtgggcca 1560 agtgggcagg gggctgaggc cgcccggggc tgggagacag ccatccgcca ggccctgatg 1620 cccgtcatcc ttcaggacgc acccagcgcc ccaggccacg

cgccccacag acaagcttct 1680 ctgagcatct ctgtgtccaa cagtcagatc caagagaatg tggacattgc cactgtctac 1740 cagatcttcc ctgacgaagt gctgggctca gggcagtttg gagtggtcta tggaggaaaa 1800 caccggaaga caggccggga cgtggcagtt aaggtcattg acaaactgcg cttccctacc 1860 aagcaggaga gccagctccg gaatgaagtg gccattctgc agagcctgcg gcatcccggg 1920 atcgtgaacc tggagtgcat gttcgagacg cctgagaaag tgtttgtggt gatggagaag 1980 ctgcatgggg acatgttgga gatgatcctg tccagtgaga agggccggct gcctgagcgc 2040 ctcaccaagt tcctcatcac ccagatcctg gtggctttga gacaccttca cttcaagaac 2100 attgtccact gtgacttgaa accagaaaac gtgttgctgg catcagcaga cccatttcct 2160 caggtgaagc tgtgtgactt tggctttgct cgcatcatcg gcgagaagtc gttccgccgc 2220 tcagtggtgg gcacgccggc ctacctggca cccgaggtgc tgctcaacca gggctacaac 2280 cgctcgctgg acatgtggtc agtgggcgtg atcatgtacg tcagcctcag cggcaccttc 2340 cctttcaacg aggatgagga catcaatgac cagatccaga acgccgcctt catgtacccg 2400 gccagcccct ggagccacat ctcagctgga gccattgacc tcatcaacaa cctgctgcag 2460 gtgaagatgc gcaaacgcta cagcgtggac aaatctctca gccacccctg gttacaggag 2520 taccagacgt ggctggacct ccgagagctg gaggggaaga tgggagagcg atacatcacg 2580 catgagagtg acgacgcgcg ctgggagcag tttgcagcag agcatccgct gcctgggtct 2640 gggctgccca cggacaggga tctcggtggg gcctgtccac cacaggacca cgacatgcag 2700 gggctggcgg agcgcatcag tgttctctga ggtcctgtgc cctcgtccag ctgctgccct 2760 ccacagcggt tcttcacagg atcccagcaa tgaactgttc tagggaaagt ggcttcctgc 2820 ccaaactgga tgggacacgt ggggagtggg gtggggggag ctatttccaa ggcccctccc 2880 tgtttcccca gcaattaaaa cggactcatc tctgccccat ggccttgatc tcaaaaaaaa 2940 aaaaaaaaaa aaaaaaaccg cggtcgcaag ctta 2974 24 3648 DNA Homo sapiens misc_feature Incyte ID No 1551456CB1 24 gggtacgagc cggatcacta ttacggcgca gtgtgctgga aagtgctcca agagatctta 60 agctggaggc accaggtctg aattccagac tcctccccac cacccacact tcacctccaa 120 ctggagcatg accacagacc cattcaggga ggctggcgga ctcttcatcc tggacagtcc 180 cttactgtat gtcaaagctg agaatgaagc ggagagcatc agacagagga gctggggaaa 240 cgtcggccag ggccaaggct ctaggaagtg ggatttctgg aaataatgca aagagagctg 300 gaccattcat ccttggtccc cgtctgggca actcaccggt gccaagcata gtgcagtgtt 360 tggcgaggaa agatggcacg gatgacttct atcagctgaa gatcctgacc ctggaggaga 420 ggggggacca aggcatagag agccaggaag agcggcaggg caagatgctg ctgcacaccg 480 agtactcact gctgtctctc ctgcacacgc aggatggcgt ggtgcaccac cacggcctct 540 tccaggaccg cacctgtgaa atcgttgagg acacagaatc cagccggatg gttaagaaga 600 tgaagaagcg catctgcctc gtcctggact gcctctgtgc tcatgacttc agcgataaga 660 ccgctgacct catcaacctg cagcactacg tcatcaagga gaagaggctc agcgagaggg 720 agactgtggt aatcttctac gacgtggtcc gcgtggtgga ggccctgcac cagaaaaata 780 tcgtgcacag agacctgaag ctggggaaca tggtgctcaa caagaggaca catcggataa 840 ccatcaccaa cttctgcctc gggaagcatc tggtgagcga gggggacctg ctgaaggacc 900 agagagggag ccctgcctac atcagtcccg acgtgctcag cggccggccg taccgtggca 960 agcccagtga catgtgggcc ctgggcgtgg tgctcttcac catgctgtat ggccagttcc 1020 ccttctacga cagcatcccg caggagctct tccgcaagat caaggctgcc gagtatacca 1080 ttcctgagga tggacgggtt tctgagaaca ccgtgtgtct catccggaag ctgctggtcc 1140 ttgaccccca gcagcgcctg gccgccgccg acgtcctgga ggccctcagt gccatcattg 1200 catcatggca gtccctgtca tctctgagtg ggcctttgca agtggttcct gacattgatg 1260 accaaatgag caatgcggat agctcccagg aggcgaaggt gacggaggag tgctcccagt 1320 acgagtttga gaactacatg cgtcagcagc tgctgctggc cgaggagaag agctccatcc 1380 atgacgcccg gagctgggta cccaagcggc agttcggcag cgcaccaccg gtgcgacggc 1440 tgggccacga cgcacagccc atgacctcct tggacacggc catcctggcg cagcgctacc 1500 tgcggaaata acagcctcag ccggggccac cagcactgct gccacttctt ccagccccag 1560 ccaaaggcgt ggctgtcagg gctgggccct gtagtgctgg actctcccgg gccacaatag 1620 ggacagggca gggacaggga cagcccaggt cacacgtggg gtcagcagag gtaccacgaa 1680 gctacctttt gggatgattg ctcgattgtt tggtttttaa atctgagaag cctagataac 1740 taatctgctt ttaatcacga tgttttaatc tacctctgtc tctttaacca tgctgtctct 1800 ggactgagca agagggagga gggagcctgc tcaccccact ccagggcctt ccccagcggc 1860 caccaactga cctggggcgc tgctccccac agtccaaata agctgaaagt gcagctcgct 1920 gcaggcccca gagcgagctt cccctcctcc ctgctctccc aggcccctgc cacagcctct 1980 ttccgtccct ctctttctga tccaggcccc tcagtccaag ctttggaaaa ccttcacctc 2040 atcttaaacc gaactcaaat atatttattt ttttaccata ccaacttctc tcccatctct 2100 aggtggctca gtccatggcc actccctgcc cccagcctgg ctggacagca aggaatccac 2160 agcccacacg tgagctccct cctcaccccc aggcagggaa gcccctcctg ccagtccctg 2220 tcccctttca gcccaccagt ccctctctgc tgccggtgat gggaggcctt tctagacctg 2280 gctctttctc tcccgtctca gtggcttctc tgaggtgctg tacacgcgcg ttaacctgtt 2340 cccttctcta tccttccccg tggtactgag ctcacgtgga ctcccagtgc gaaggggccc 2400 atgggttggg ctgcaggcct ggccgtgagc gggggctgcc tgcacgctcc cctagcctac 2460 tcttgtgttt aggggatggt gggaacatat cccagtgccc ttgcctcata atagatgtgg 2520 tgactctccc ggtagaccct agcaagggtc ctccatggtg gtgagggact caggagaatt 2580 gtagggattg ggggaccctg cctgcctggc ttgagaacag ccctgctgcc cttttgagcc 2640 gagattttga agtggatgcc cgtcttgcca gaaatgctgt tctcaccaga atgccccctc 2700 cccttgccct tactggactt ggccctgcct gatgccaagc aaagaccctt ccccagaggc 2760 ctacccccca tatgtcctca gagaggctga gtgtcccctc caggcagtca tgggccctga 2820 ggcccctcct gcctggccct gctccccagt ggggaggtga ctgtgtttcc cagagtgtga 2880 gccgctctcc tccccctaaa aagctgactc actgtgagtg accttgggca agttcccaaa 2940 cctccttgtg cctcagtttc cccatctgga aaaaatgggg ccacctcttg ccagcagtag 3000 cagggctgcc cacgcccctt tctccccatg ccccatccag cacttgggcg actcatgcct 3060 ctgcctcagt gggcctgtgg gagcctactg gagcccagca cttactcccc ctgagcagcg 3120 agcctgcgtc tgtctcagct gtccagcgct gagggccagg gtcttgtgct gtggggctgg 3180 gggatgccct cttttctata tttatttcat agaaagtctc ctgcgggagc ggaaatgcag 3240 tccggcctag ggctcccagc ccttgactgt cctcctgtga gggcctgaag ctgggccagg 3300 gcccgtcgca gcggagcccc ctctcagcag cccaccgggt ccctccaggc tgctgcccgt 3360 gcgtggtctt tctcctcctt ttcaaagcaa tagccgccgg gtctgcaaag ccctgtcaga 3420 cagactgggc ccttccaagg tcaagccatg tgtctgatga cattcctggt gaagcaaagg 3480 agaggaggat gggtcagccc tcactgggtg tcacacactg agagaagtcc tattgtaaag 3540 aaacggaaaa agtcacaaaa aagtttgtat aaagacatat ttttgtacta catggggact 3600 cttcctgcat gtcagcaata aaacttcctg atctggaaaa aaaaaaaa 3648 25 4719 DNA Homo sapiens misc_feature Incyte ID No 2589355CB1 25 gcggagcggc cgtcgcccaa gccaagccgc gctgccaacc ctcccgcccg cccgcgctcc 60 tgtccgccgt gtctagcagc ggggcccagc atggtcatgg cggatggccc gaggcacttg 120 cagcgcgggc cggtccgggt ggggttctac gacatcgagg gcacgctggg caagtggcaa 180 cttcgctgtg gtgaagctgg ggcggcaccg gatcaccaag acggaggtgg caataaaaat 240 aatcgataag tctcagccgt ggatgcatgt gaaccttgag aaaatctacc gagaagtaca 300 aataatgaaa atgttagacc accctcacat aatcaaactt tatcaggtaa tggagaccaa 360 aagtatgttg taccttgtga cagaatatgc caaaaatgga gaaatttttg actatcttgc 420 taatcatggc cggttaaatg agtctgaagc caggcgaaaa ttctggcaaa tcctgtctgc 480 tgttgattat tgtcatggtc ggaagattgt gcaccgtgac ctcaaagctg aaaatctcct 540 gctggataac aacatgaata tcaaaatagc agatttcggt tttggaaatt tctttaaaag 600 tggtgaactg ctggcaacat ggtgtggcag ccccccttat gcagccccag aagtctttga 660 agggcagcag tatgaaggac cacagctgga catctggagt atgggagttg ttctttatgt 720 ccttgtctgt ggagctctgc cctttgatgg accgactctt ccaattttga ggcagagggt 780 tctggaagga agattccgga ttccgtattt catgtcagaa gattgcgagc accttatccg 840 aaggatgttg gtcctagacc catccaaacg gctaaccata gcccaaatca aggagcataa 900 atggatgctc atagaagttc ctgtccagag acctgttctc tatccacaag agcaagaaaa 960 tgagccatcc atcggagagt ttaatgagca ggttctgcga ctgatgcaca gccttggaat 1020 agatcagcag aaaaccattg agtctttgca gaacaagagc tataaccact ttgctgccat 1080 ttatttcttg ttggtggagc gcctgaaatc acatcggagc agtttcccag tggagcagag 1140 acttgatggc cgccagcgtc ggcctagcac cattgctgag caaacagttg ccaaggcaca 1200 gactgtgggg ctcccagtga ccatgcattc accgaacatg aggctgctgc gatctgccct 1260 cctcccccag gcatccaacg tggaggcctt ttcatttcca gcatctggct gtcaggcgga 1320 agctgcattc atggaagaag agtgtgtgga cactccaaag gtcaatggct gtctgcttga 1380 ccctgtgcct cctgtcctgg tgcggaaggg atgccagtca ctgcccagca acatgatgga 1440 gacctccatt gacgaagggc tggagacaga aggagaggcc gaggaagacc ccgctcatgc 1500 ctttgaggca tttcagtcca cacgcagcgg gcagagacgg cacactctgt cagaagtgac 1560 caatcaactg gtcgtgatgc ctggggcagg gaaaattttc tccatgaatg acagcccctc 1620 ccttgacagt gtggactctg agtatgatat ggggtctgtt cagagggacc tgaactttct 1680 ggaagacaac ccttccctta aggacatcat gttagccaat cagccttcac cccgcatgac 1740 atctcccttc ataagcctga gacctaccaa cccagccatg caggctctga gctcccagaa 1800 acgagaggtc cacaacaggt ctccagtgag cttcagagag ggccgcagag catcagatac 1860 ctccctcacc cagggaattg tagcatttag acaacatctt cagaatctgg ctagaaccaa 1920 aggaattcta gagttgaaca aagtgcagtt gttgtatgaa caaataggac cggaggcaga 1980 ccctaacctg gcgccggcgg ctcctcagct ccaggacctt gctagcagct gccctcagga 2040 agaagtttct cagcagcagg aaagcgtctc cactctccct gccagcgtgc atccccagct 2100 gtccccacgg cagagcctgg agacccagta cctgcagcac agactccaga agcccagcct 2160 tctgtcaaag gcccagaaca cctgtcagct ttattgcaaa gaaccaccgc ggagccttga 2220 gcagcagctg caggaacata ggctccagca gaagcgactc tttcttcaga agcagtctca 2280 actgcaggcc tattttaatc agatgcagat agcagagagc tcctacccac agccaagtca 2340 gcagctgccc cttccccgcc aggagactcc accgccttct cagcaggccc caccgttcag 2400 cctgacccag cccctgagcc ccgtcctgga gccttcctcc gagcagatgc aatacagccc 2460 tttcctcagc cagtaccaag agatgcagct tcagcccctg ccctccactt ccggtccccg 2520 ggctgctcct cctctgccca cgcagctaca gcagcagcag ccgccaccgc caccaccccc 2580 tccaccacca cgacagccag gagctgcccc agccccctta cagttctcct atcagacttg 2640 tgagctgcca agcgctgctt cccctgcgcc agactatccc actccctgtc agtatcctgt 2700 ggatggagcc cagcagagcg acctaacggg gccagactgt cccagaagcc caggactgca 2760 agaggccccc tccagctacg acccactagc cctctctgag ctacctggac tctttgattg 2820 tgaaatgcta gacgctgtgg atccacaaca caacgggtat gtcctggtga attagtctca 2880 gcacaggaat tgaggtgggt caggtgaagg aagagtgtat gttcctattt ttattccagc 2940 cttttaaatt taaagcttat tttcttgccc tctccctaac ggggagaaat cgagccaccc 3000 aactggaatc agagggtctg gctggggtgg atgttgcttc ctcctggttc tgccccacca 3060 caaagttttc tgtggcaagt gctggaacat agttgtaggc tgaggctcct gcccttcggt 3120 cgagtggagc aagctctcga gggcagcact gacaaatgtg ttcctaagaa gacattcaga 3180 cccaggtctt atgcaggatt acatccgttt attatcaagg gcaaccttgg tgaaagcaga 3240 aagggtgtgt gctattgcat atatatgggg gaaaaggcaa tatatttttc actgaagctg 3300 agcaaccaca tattgctaca aggcaaatca agaagacatc aggaaatcag atgcacagga 3360 aataaaggaa agctgtgctt tgtcattgaa tcctaagttc ttagctgctg atgcaagttg 3420 tcccccaagg ccatcacaaa gcagtggggc atgagctgtg tttcaggggc cactaaataa 3480 cagctggtac tgaccccaga aaccgccttc atctccattc ggaagcaggt gacacacccc 3540 ttcagaaggt gccctgggtt gccgagtgtc agaatatact caggactcca gaggtgtcac 3600 acgtggaact gacaggagac ccgccaccgt ggaggcaggg ggcaagaaac tcaagaacgc 3660 atcaagagca ccagccctgg gccagggaag acaggctctt cctgcagttt ctcgtggaca 3720 ctgctggctt gcgggcagtc ggtctccagg gtacctgttg tctcttttcc gatgtaataa 3780 ctactttgac cttacactat atgttgctag tagtttattg agctttgtat atttggacag 3840 tttcatatag ggcttagaga ttttaaggac atgataaatg aacttttctg tcccatgtga 3900 agtggtagtg cggtgccttt cccccagatc atgctttaat tctttctttt ctgtagaaac 3960 caacagtttc catttatgtc aatgctaaat ccaaagtcac ttcagagttt gttttccacc 4020 atgtgggaat cagcattctt aatttcgtta aagttttgac ttgtaatgaa atgttcaagt 4080 attacagcaa tattcaaaga aagaaccaca gatgtgttaa ccatttaagc agatcatctg 4140 ccaaacatta tattactaat aaaacttaac caacacttac aattcagtca tcaaagtaag 4200 taaaaattag atgctacagc tagctaactg tatccctaga aatgatgaat aatttgccat 4260 ttggacagtt aacatccagg tgttacaaag tcagtgttaa ttctaaagat gatcatttct 4320 gccctttaga atggcttgtc ccatcagcag atgaatgtgt taagcacaaa gcatcttcct 4380 taaagcacaa agagagggac taactgatgc tgcatctaga aaacaccttt aagttgcctt 4440 tcctctttgt agttagcgtt caggcaggtg acgtgtggaa agtctagggg gttccattct 4500 ggccatgcga gcccagctcc taccaacgtc ggtaacttga gcagtccctg ttgctggcca 4560 gagactgcct ggtcgccagc gctcaacatg ggtgccagga tgcttcgcag aggcactgtg 4620 ctcacggttg gacttggtgt cagtgggaaa gggcagtgtg gggactgtca tttttgtgat 4680 ttataacaca cgtgaaaatc aggaagaatg aataagctt 4719 26 1651 DNA Homo sapiens misc_feature Incyte ID No 4357117CB1 26 atgaggattg tttgtttagt gaaaaaccaa cagcccctgg gagccaccat caagcgccac 60 gagatgacag gggacatctt ggtggccagg atcatccacg gtgggctggc ggagagaagt 120 ggtttgctat atgctggaga caaactggta gaagtgaatg gagtttcagt tgagggactg 180 gaccctgaac aagtgatcca tattctggcc atgtctcgag gcacaatcat gttcaaggtg 240 gttccagtct ctgaccctcc tgtgaatagc cagcagatgg tatacgtccg tgccatgact 300 gagtactggc cccaggagga tcccgacatc ccctgcatgg acgctggatt gcctttccag 360 aagggggaca tcctccagat tgtggaccag aatgatgccc tctggtggca ggcccgaaaa 420 atctcagacc ctgctacctg cgctgggctt gtcccttcta accaccttct gaagaggaag 480 caacgggaat tctggtggtc tcagccgtac cagcctcaca cctgcctcaa gtcaacctca 540 gacaaggagg agtttgttgg ctacggtcag aagttcttta taggtaggtt cagcccgctg 600 catgccagtg tgtgctgcac cggcagctgc tacagtgcag tgggtgcccc ttacgaggag 660 gtggtgaggt accagcgacg cccttcagac aagtaccgcc tcatagtgct cataggaccc 720 tctggtgttg gagtaaatga gctcagaaga caacttattg aatttaatcc cagccatttt 780 caaagtgctg tgccacacac tactcgtact aaaaagagtt acgaaatgaa tgggcgtgag 840 tatcactatg tgtccaagga aacatttgaa aacctcatat atagtcacag gatgctggag 900 tatggtgagt acaaaggcca cctgtatggc actagtgtgg atgctgttca aacagtcctt 960 gtcgaaggaa agatctgtgt catggaccta gagcctcagg atattcaagg ggttcgaacc 1020 catgaactga agccctatgt catatttata aagccatcga atatgaggtg tatgaaacaa 1080 tctcggaaaa atgccaaggt tattactgac tactatgtgg acatgaagtt caaggatgaa 1140 gacctacaag agatggaaaa tttagcccaa agaatggaaa ctcagtttgg ccaatttttt 1200 gatcatgtga ttgtgaatga cagcttgcac gatgcatgtg cccagttgtt gtctgccata 1260 cagaaggctc aggaggagcc tcagtgggta ccagcaacat ggatttcctc agatactgag 1320 tctcaatgag acttcttgtt taatgctgga gttttaacac tgtacccttg atacagcgat 1380 ccatagttgc aatctaaaac aacagtattt gacccatttt aatgtgtaca actttaaaag 1440 tgcagcaatt tattaattaa tcttatttga aaaaaatttt tattgtatgg ttatgtggtt 1500 acctatttta acttaatttt ttttccttta cctcatatgc agctgtggta gaaatatgaa 1560 taatgttaag tcactgagta tgagaacctt tcgcagattt cacatgatct ttttaagatt 1620 taaataaaga gctttcctaa ataaaaaaaa a 1651 27 3141 DNA Homo sapiens misc_feature Incyte ID No 5511992CB1 27 atggagccct ccagagcgct tctcggctgc ctagcgagcg ccgccgctgc cgccccgccg 60 ggggaggatg gagcaggggc cggggccgag gaggaggagg aggaggagga ggaggcggcg 120 gcggcggtgg gccccgggga gctgggctgc gacgcgccgc tgccctactg gacggccgtg 180 ttcgagtacg aggcggcggg cgaggacgag ctgaccctgc ggctgggcga cgtggtggag 240 gtgctgtcca aggactcgca ggtgtccggc gacgagggct ggtggaccgg gcagctgaac 300 cagcgggtgg gcatcttccc cagcaactac gtgaccccgc gcagcgcctt ctccagccgc 360 tgccagcccg gcggcgagga agaaattgat tttgcggagc tcaccttgga agagattatt 420 ggcatcgggg gctttgggaa ggtctatcgt gctttctgga taggggatga ggttgctgtg 480 aaagcagctc gccacgaccc tgatgaggac atcagccaga ccatagagaa tgttcgccaa 540 gaggccaagc tcttcgccat gctgaagcac cccaacatca ttgccctaag aggggtatgt 600 ctgaaggagc ccaacctctg cttggtcatg gagtttgctc gtggaggacc tttgaataga 660 gtgttatctg ggaaaaggat tcccccagac atcctggtga attgggctgt gcagattgcc 720 agagggatga actacttact tgatgaggca attgttccca tcatccaccg cgaccttaag 780 tccagcaaca tattgatcct ccagaaggtg gagaatggag acctgagcaa caagattctg 840 aagatcactg attttggcct ggctcgggaa tggcaccgaa ccaccaagat gagtgcggca 900 gggacgtatg cttggatggc acccgaagtc atccgggcct ccatgttttc caaaggcagt 960 gatgtgtgga gctatggggt gctactttgg gagttgctga ctggtgaggt gccctttcga 1020 ggcattgatg gcttagcagt cgcttatgga gtggccatga acaaactcgc ccttcctatt 1080 ccttctacgt gcccagaacc ttttgccaaa ctcatggaag actgctggaa tcctgatccc 1140 cactcacgac catctttcac gaatatcctg gaccagctaa ccaccataga ggagtctggt 1200 ttctttgaaa tgcccaagga ctccttccac tgcctgcagg acaactggaa acacgagatt 1260 caggagatgt ttgaccaact cagggccaaa gaaaaggaac ttcgcacctg ggaggaggag 1320 ctgacgcggg ctgcactgca gcagaagaac caggaggaac tgctgcggcg tcgggagcag 1380 gagctggccg agcgggagat tgacatcctg gaacgggagc tcaacatcat catccaccag 1440 ctgtgccagg agaagccccg ggtgaagaaa cgcaagggca agttcaggaa gagccggctg 1500 aagctcaagg atggcaaccg catcagcctc ccttctggtt tccagcacaa gttcacggtg 1560 caggcctccc ctaccatgga taaaaggaag agtcttatca acagccgctc cagtcctcct 1620 gcaagcccca ccatcattcc tcgccttcga gccatccagt gtgagactgt tagcaaaacc 1680 tggggcagga gctcagtcgt cccaaaggag gaaggggagg aggaggagaa gagggcccca 1740 aagaagaagg gacggacgtg ggggccaggg acgcttggtc agaaggagct tgcctcggga 1800 gatgaaagcc tcaagtccct ggtagatgga tataagcagt ggtcgtccag tgcccccaac 1860 ctggtgaagg gcccaaggag tagcccggcc ctgccagggt tcaccagcct tatggagatg 1920 ggtaagttca cagaggatga ggacagtgaa ggcccaggga gtggagagag tcgcctacag 1980 cattcaccca gccagtccta cctctgtatc ccattccctc gtggagagga tggcgatggc 2040 ccctccagtg atggaatcca tgaggagccc accccagtca actcggccac gagtacccct 2100 cagctgacgc caaccaacag cctcaagcgg ggcggtgccc accaccgccg ctgcgaggtg 2160 gctctgctcg gctgtggggc tgttctggca gccacaggcc tagggtttga cttgctggaa 2220 gctggcaagt gccagctgct tcccctggag gagcctgagc caccagcccg ggaggagaag 2280 aaaagacggg agggtctttt tcagaggtcc agccgtcctc gtcggagcac cagcccccca 2340 tcccgaaagc ttttcaagaa ggaggagccc atgctgttgc taggagaccc ctctgcctcc 2400 ctgacgctgc tctccctctc ctccatctcc gagtgcaact ccacacgctc cctgctgcgc 2460 tccgacagcg atgaaattgt cgtgtatgag atgccagtca gcccagtcga ggcccctccc 2520 ctgagtccat gtacccacaa ccccctggtc aatgtccgag tagagcgctt caaacgagat 2580 cctaaccaat ctctgactcc cacccatgtc accctcacca ccccctcgca gcccagcagt 2640 caccggcgga ctccttctga tggggccctt cccagtccca gccgagaccc aggtgaattc 2700 ccccgtctcc ctgaccccaa tgtggtcttc cccccaaccc caaggcgctg gaacactcag 2760 caggactcta ccttggagag acccaagact ctggagtttc tgcctcggcc gcgtccttct 2820 gccaaccggc aacggctgga cccttggtgg tttgtgtccc ccagccatgc ccgcagcacc 2880 tccccagcca acagctccag cacagagacg cccagcaacc tggactcctg ctttgctagc 2940 agtagcagca ctgtagagga gcggcctgga cttccagccc tgctcccgtt ccaggcaggg 3000 ccgctgcccc cgactgagcg gacgctcctg gacctggatg cagaggggca gagtcaggac 3060 agcaccgtgc cgctgtgcag agcggaactg aacacacaca ggcctgcccc ttatgagatc 3120 cagcaggagt tctggtctta g 3141 28 1244 DNA Homo sapiens misc_feature Incyte ID No 7474560CB1 28 gtcttattgc caagataatt acaactggag aattggcccc acaggaaaca acaattacag 60 agataaaaca aaaattgatg caaatacctg atgaagaggg cattgttatt gatggatttc 120 caagagatgt tgcccaggct ctatcttttg aggaccaaat

ctgtaccccc gatttggtgg 180 tattcctggc ttgtgctaat cagagactca aagaaagatt actgaagcgt gcagaacagc 240 agggccgacc agacgacaat gtaaaagcta cccaaaggag actaatgaac ttcaagcaga 300 atgctgctcc attggttaaa tacttccagg aaaaggggct catcatgaca tttgatgccg 360 accgcgatga ggatgaggtg ttctatgaca tcagcatggc agttgacaac aagttatttc 420 caaacaaaga ggctgcagca ggttcaagtg accttgatcc ttcgatgata ttggacactg 480 gagagatcat tgatacagga tctgattatg aagatcaggg tgatgaccag ttaaatgtat 540 ttggagagga cactatggga ggtttcatgg aagatttgag aaagtgtaaa attattttca 600 taattggtgg tcctggctct ggcaaaggca cacagtgtga aaagctggtg gaaaaatatg 660 gatttacaca tctctcaact ggcgagctcc tgcgtgagga actggcatca gaatctgaaa 720 gaagcaaatt gatcagagac attatggaac gtggagacct ggtgccctca ggcatcgttt 780 tggagctcct gaaggaggcc atggtggcca gcctcgggga caccaggggc ttcctgattg 840 acggctatcc tcgggaggtg aagcaagggg aagagttcgg acgcaggatt ggagacccac 900 agttggtgat ctgtatggac tgctcggcag acaccatgac caaccgcctt ctccaaagga 960 gccggagcag cctgcctgtg gacgacacca ccaagaccat cgccaagcgc ctagaagcct 1020 actaccgagc gtccatcccc gtgatcgcct actacgagac aaaaacacag ctacacaaga 1080 taaatgcaga gggaacacca gaggacgttt ttcttcaact ctgcacagct attgactcta 1140 ttattttctg aaggcaaaaa tgcatgttag aatggaaaca gaaaaacatt aaaaagttca 1200 ttccgttaac acaatgtttc aagttaaacc ttttgtgtca ccgc 1244 29 1661 DNA Homo sapiens misc_feature Incyte ID No 7474602CB1 29 gcggcggcgg cgagagcgaa agaggaaact gcagaggagg aagctgcgcc gcagcccgag 60 ccgcccggca tccccgccgc ctctgcgccc gcgccgcgcc cccggcgccc cctccccagc 120 gcgcccccgg ccgctcctcc gcgccgcgct cgtcggccat ggcccgggag aacggcgaga 180 gcagctcctc ctggaaaaag caagctgaag acatcaagaa gatcttcgag ttcaaagaga 240 ccctcggaac cggggccttt tccgaagtgg ttttagctga agagaaggca actggcaagc 300 tctttgctgt gaagtgtatc cctaagaagg cgctgaaggg caaggaaagc agcatagaga 360 atgagatagc cgtcctgaga aagattaagc atgaaaatat tgttgccctg gaagacattt 420 atgaaagccc aaatcacctg tacttggtca tgcagctggt gtccggtgga gagctgtttg 480 accggatagt ggagaagggg ttttatacag agaaggatgc cagcactctg atccgccaag 540 tcttggacgc cgtgtactat ctccacagaa tgggcatcgt ccacagagac ctcaagcccg 600 aaaatctctt gtactacagt caagatgagg agtccaaaat aatgatcagt gactttggat 660 tgtcaaaaat ggagggcaaa ggagatgtga tgtccactgc ctgtggaact ccaggctatg 720 tcgctcctga agtcctcgcc cagaaacctt acagcaaagc cgttgactgc tggtccatcg 780 gagtgattgc ctacatcttg ctctgcggct accctccttt ttatgatgaa aatgactcca 840 agctctttga gcagatcctc aaggcggaat atgagtttga ctctccctac tgggatgaca 900 tctccgactc tgcaaaagac ttcattcgga acctgatgga gaaggacccg aataaaagat 960 acacgtgtga gcaggcagct cggcacccat ggatcgctgg tgacacagcc ctcaacaaaa 1020 acatccacga gtccgtcagc gcccagatcc ggaaaaactt tgccaagagc aaatggagac 1080 aagcatttaa tgccacggcc gtcgtgagac atatgagaaa actacacctc ggcagcagcc 1140 tggacagttc aaatgcaagt gtttcgagca gcctcagttt ggccagccaa aaagactgtg 1200 cgtctggcac cttccacgct ctgtagtttc atttcttctt cgtcgggggt ctcaggagtt 1260 ggagccgagc ggagacccag gcccaccact gtgacggcag tgcactctgg aagcaagtga 1320 ctggccctgg aggtggggcc cggggtcggg gctggggaag gggagcccca gggtcgccag 1380 agccgcgagc cactccagcg agaccccacc ttgcatggtg ccccttcctg cataggactg 1440 gaagaccgaa gtttttttat ggccatattt tctactgcaa ttctgaagtg ttcatttctc 1500 acaaactgta ctgactcgag gggcgctgat ttcataggat ctggtgctgt atatacgaat 1560 cttgcaaagc tctaactgaa cggaccttct tattcctctc tcctaacacc atcgtttcca 1620 ctcttctcag tgtaggtaac cgtctatggt gtgtctttca t 1661 30 912 DNA Homo sapiens misc_feature Incyte ID No 7475509CB1 30 cggacggtgg gctcggtccc ggcgctgggc tgaggggagg ggttgtctta aaagtctctc 60 cttccccctg taggggcggc cggcgagtcc cagtgagagc ggagggtgcc agaggtaggg 120 ggccgagaaa caaagttccc ggggcttcct ccggggccgc ggtcggggct gcgcgtttga 180 ccgcccccct cctcgcgaag gcaatggctt ccaaactcct gcgcgcggtc atcctcgggc 240 cgcccggctc gggcaagggc accgtgtgcc agaggatcgc ccagaacttt ggtctccagc 300 atctctccag cggccacttc ttgcgggaga acatcaaggc cagcaccgaa gttggtgaga 360 tggcaaagca gtatatagag aaaagtcttt tggttccaga ccatgtgatc acacgcctaa 420 tgatgtccga gttggagaac aggcgtggcc agcactggct ccttgatggt tttcctagga 480 cattaggaca agccgaagcc ctggacaaaa tctgtgaagt ggatctagtg atcagtttga 540 atattccatt tgaaacactt aaagatcgtc tcagccgccg ttggattcac cctcctagcg 600 gaagggtata taacctggac ttcaatccac ctcatgtaca tggtattgat gacgtcactg 660 gtgaaccgtt agtccagcag gaggatgata aacccgaagc agttgctgcc aggctaagac 720 agtacaaaga cgtggcaaag ccagtcattg aattatacaa gagccgagga gtgctccacc 780 aatttttccg gaaccggaga cgaacaaaaa tctggcctta cgtttacaca acttttctca 840 acaagatcac acctattcag tccaaggaag cattttgacc tgcccatgga gaccggaatg 900 ggcctcactc ct 912 31 2858 DNA Homo sapiens misc_feature Incyte ID No 7475491CB1 31 ggtacgagcc ggatcacttg tacggcgcag tgtgctggac agactctctc cttcccctcc 60 ccttctacct cctcctcggc cagctcaggt tgcagcttct ctggggaact gctcaccttt 120 ccggagcagg ggaagctgcc ccgtgcccgg gagggagcgg gcgcaccgcg gcccccagga 180 cacgcgctgt gagtccgcgg gcggtgcgcc tgggaggaag ggggaggtcg gaggaggggg 240 caccgcggcg ccgggtataa ggagcaaagg acccggctgc ccagtccctc atgatcatga 300 acaagatgaa gaactttaag cgccgtttct ccctgtcagt gccccgcact gagaccattg 360 aagaatcctt ggctgaattc acggagcaat tcaaccagct ccacaaccgg cggaatgaga 420 acttgcagct cggtcctctt ggcagagacc ccccgcagga gtgcagcacc ttctccccaa 480 cagacagcgg ggaggagccg gggcagctct cccctggcgt gcagttccag cggcggcaga 540 accagcgccg cttctccatg gaggtgaggg cctctggagc tctgccccgg caggtggcag 600 gatgcacgca caagggtgtg cacaggaggg cagctgcctt acagccagac tttgacgtca 660 gcaagaggct ctctctgccc atggatatcc gcctgcccca ggaattccta cagaagctac 720 agatggagag cccagatctg cccaagccgc tcagccgcat gtcccgccgg gcctccctgt 780 cagacattgg ctttgggaaa ctggaaacat acgtgaaact ggacaaactg ggagagggca 840 cctatgccac agtcttcaaa gggcgcagca aactgacgga gaaccttgtg gccctgaaag 900 agatccggct ggagcacgag gagggagcgc cctgcactgc catccgagag gtgtctctgc 960 tgaagaacct gaagcacgcc aatattgtga ccctgcatga cctcatccac acagatcggt 1020 ccctcaccct ggtgtttgag tacctggaca gtgacctgaa gcagtatctg gaccactgtg 1080 ggaacctcat gagcatgcac aacgtcaaga ttttcatgtt ccagctgctc cggggcctcg 1140 cctactgtca ccaccgcaag atcctgcacc gggacctgaa gccccagaac ctgctcatca 1200 acgagagggg ggagctgaag ctggccgact ttggactggc cagggccaag tcagtgccca 1260 caaagactta ctccaatgag gtggtgaccc tgtggtacag gccccccgat gtgctgctgg 1320 gatccacaga gtactccacc cccattgata tgtggggcgt gggctgcatc cactacgaga 1380 tggccacagg gaggcccctc ttcccgggct ccacagtcaa ggaggagctg cacctcatct 1440 ttcgcctcct cgggaccccc acagaagaga cgtggcccgg cgtgaccgcc ttctctgagt 1500 tccgcaccta cagcttcccc tgctacctcc cgcagccgct catcaaccac gcgcccaggt 1560 tggatacgga tggcatccac ctcctgagca gcctgctcct gtatgaatcc aagagtcgca 1620 tgtcagcaga ggctgccctg agtcactcct acttccggtc tctgggagag cgtgtgcacc 1680 agcttgaaga cactgcctcc atcttctccc tgaaggagat ccagctccag aaggacccag 1740 gctaccgagg cttggccttc cagcagccag gacgagggaa gaacaggcgg cagagcatct 1800 tctgagccac gcccaccttg ctgtggccaa gggacaagag atcacatgga gcacaaattc 1860 gggtaggatg gagcctgtgt ggccctcgga ggactgaaga acgagggctg acagccagcc 1920 tggaagaccg cttggcagcc cttctggcca cggctgtttc ttctttgtgc ttcccgtgtg 1980 cctccccagt agccctcacc tgcataccaa cccctccttt acccacgttg gggctggcat 2040 aagctgcttc cctgagagga catgaggggg gggcggtcct cgtaccctct cccaccctgg 2100 tgtttgggca cctgcgtggg atgcacacgg atgacagaat caaggcgcca ggatgggcac 2160 tctgccctgg atacaggctc taccctcctc ccccaggacc tgcctagtgc cagtttggta 2220 gtcccccttt ctggcccctt ggagcccaca cacgtttcat ctttttcccc tctgagagca 2280 agaagagaca tggcatgttc tctgggaccc tggaatccta ggtacccaca tgtgtgccaa 2340 agcctacccc acctggcagg tgtcccacag caacagaagg aatagtagtc cccactcttt 2400 ccatcagccc taccctaccc tcattccccg acaccctctg gcttgaacca tggctgagca 2460 gtgccggcat acgctttgct ggcatgcttg gatgcccagc tgtgtccaga ggtggcctgg 2520 gaccgccagt tgcacgcctg ccacctcagc cagcccccgc ccagctcatc agtctgaatg 2580 gagttgcctt aaattggcag gtggtaccgt actcactgcc cttggagctg tgaccggctc 2640 ctgcctgtcc accccttccc gaggtggctc ctgcttacct tatcatccca gggctctgat 2700 tagccaggcc tggtcagggt cctggggacg gcacccagat atgcagagtc accctgacac 2760 tggtgccagg ctgacctcag ctcccgaagg ctcgcacagc ctccccatcc ttccttccca 2820 gcccttgtgg ctctgtccac ctgatcccaa taccagct 2858 32 2817 DNA Homo sapiens misc_feature Incyte ID No 2192119CB1 32 ttcttctatc ctgaaagctg tcctctcccc actcccgaca cactgggagc ccccccatcc 60 ccgcccctct cccgcccctc ccttgtcccc gccccttccc gccccaactc aggttctgcc 120 ccaccccgcc ccaattcacg tcctgctccg ccccgacgta ggccccgccc tggcccccgg 180 gccccgcccc cgtctgacgc aggccccgcc ccctctccgc cccgccccgg ctcgggcggc 240 cggaggaccc ggagctaagg cgcccgaacc cgcggcggcg gtggggacga tgtggttctt 300 tgcccgggac ccggtccggg actttccgtt cgagctcatc ccggagcccc cagagggcgg 360 cctgcccggg ccctgggccc tgcaccgcgg ccgcaagaag gccacaggca gccccgtgtc 420 catcttcgtc tatgatgtga agcctggcgc ggaagagcag acccaggtgg ccaaagctgc 480 cttcaagcgc ttcaaaactc tacggcaccc caacatcctg gcttacatcg atggactgga 540 gacagaaaaa tgcctccacg tcgtgacaga ggctgtgacc ccgttgggaa tatacctcaa 600 ggcgagagtg gaggctggtg gcctgaagga gctggagatc tcctgggggc tacaccagat 660 cgtgaaagcc ctcagcttcc tggtcaacga ctgcagcctc atccacaaca atgtctgcat 720 ggccgccgtg ttcgtggacc gagctggcga gtggaagctt gggggcctgg actacatgta 780 ttcggcccag ggcaacggtg ggggacctcc ccgcaagggg atccccgagc ttgagcagta 840 tgaccccccg gagttggctg acagcagtgg cagagtggtc agagagaagt ggtcagcaga 900 catgtggcgc ttgggctgcc tcatttggga agtcttcaat gggcccctac ctcgggcagc 960 agccctacgc aaccctggga agatccccaa aacgctggtg ccccattact gtgagctggt 1020 gggagcaaac cccaaggtgc gtcccaaccc agcccgcttc ctgcagaact gccgggcacc 1080 tggtggcttc atgagcaacc gctttgtaga aaccaacctc ttcctggagg agattcagat 1140 caaagagcca gccgagaagc aaaaattctt ccaggagctg agcaagagcc tggacgcatt 1200 ccctgaggat ttctgtcggc acaaggtgct gccccagctg ctgaccgcct tcgagttcgg 1260 caatgctggg gccgttgtcc tcacgcccct cttcaaggtg ggcaagttcc tgagcgctga 1320 ggagtatcag cagaagatca tccctgtggt ggtcaagatg ttctcatcca ctgaccgggc 1380 catgcgcatc cgcctcctgc agcagatgga gcagttcatc cagtaccttg acgagccaac 1440 agtcaacacc cagatcttcc cccacgtcgt acatggcttc ctggacacca accctgccat 1500 ccgggagcag acggtcaagt ccatgctgct cctggcccca aagctgaacg aggccaacct 1560 caatgtggag ctgatgaagc actttgcacg gctacaggcc aaggatgaac agggccccat 1620 ccgctgcaac accacagtct gcctgggcaa aatcggctcc tacctcagtg ctagcaccag 1680 acacagggtc cttacctctg ccttcagccg agccactagg gacccgtttg caccgtcccg 1740 ggttgcgggt gtcctgggct ttgctgccac ccacaacctc tactcaatga acgactgtgc 1800 ccagaagatc ctgcctgtgc tctgcggtct cactgtagat cctgagaaat ccgtgcgaga 1860 ccaggccttc aaggccattc ggagcttcct gtccaaattg gagtctgtgt cggaggaccc 1920 gacccagctg gaggaagtgg agaaggatgt ccatgcagcc tccagccctg gcatgggagg 1980 agccgcagct agctgggcag gctgggccgt gaccggggtc tcctcactca cctccaagct 2040 gatccgttcg cacccaacca ctgccccaac agaaaccaac attccccaaa gacccacgcc 2100 tgaaggccac tgggagacgc aggaggagga caaggacaca gcagaggaca gcagcactgc 2160 tgacagatgg gacgacgaag actggggcag cctggagcag gaggccgagt ctgtgctggc 2220 ccagcaggac gactggagca ccgggggcca agtgagccgt gctagtcagg tcagcaactc 2280 cgaccacaaa tcctccaaat ccccagagtc cgactggagc agctgggaag ctgagggctc 2340 ctgggaacag ggctggcagg agccaagctc ccaggagcca cctcctgacg gtacacggct 2400 ggccagcgag tataactggg gtggcccaga gtccagcgac aagggcgacc ccttcgctac 2460 cctgtctgca cgtcccagca cccagccgag gccagactct tggggtgagg acaactggga 2520 gggcctcgag actgacagtc gacaggtcaa ggctgagctg gcccggaaga agcgcgagga 2580 gcggcggcgg gagatggagg ccaaacgcgc cgagaggaag gtggccaagg gccccatgaa 2640 gctgggagcc cggaagctgg actgaaccgt ggcggtggcc cttcccggct gcggagagcc 2700 cgccccacag atgtatttat tgtacaaacc atgtgagccc ggccggccca gccaggccat 2760 ctcacgtgta cataatcaga gccacaataa attctatttc acaaaaaaaa aaaaaaa 2817 33 5305 DNA Homo sapiens misc_feature Incyte ID No 7474496CB1 33 atggctgggg gccgtggggc ccccgggcgc ggccgggacg agcctccgga gagctacccg 60 caacgacagg accacgagct acaggccctg gaggccatct acggcgcgga cttccaagac 120 ctgcggccgg acgcttgcgg accggtcaaa gagccccctg aaatcaattt agttttgtac 180 cctcaaggcc taactggtga agaagtatat gtaaaagtgg atttgagggt taaatgccca 240 cctacctatc cagatgtagt tcctgaaata gagttaaaaa atgccaaagg tctatcaaat 300 gaaagtgtca atttgttaaa atctcgccta gaagaactgg ccaagaaaca ctgtggggag 360 gtgatgatct ttgaactggc ttaccacgtg cagtcatttc tcagcgagca taacaagccc 420 cctcccaagt cttttcatga agaaatgctg gaaaggcggg ctcaagagga gcaacagagg 480 ctgttggagg cccaagcgga aagaagaaga gagcaggcac aacaacgtga aatcctgcat 540 gagattcaga gaaggaaaga agagataaaa gaagagaaaa aaaggaaaga aatggctaag 600 caggaacgtt tggaaattgc tagtttgtca aaccaagatc atacctctaa gaaggaccca 660 ggaggacaca gaacggctgc cattctacat ggaggctctc ctgactttgt aggaaatggt 720 aaacatcggg caaactcctc aggaaggtct aggcgagaac gtcagtattc tgtatgtaat 780 agtgaagatt ctcctggctc ttgtgaaatt ctgtatttca atatggggag tcctgatcag 840 ctcatggtgc acaaagggaa atgtattggc agtgatgaac aacttggaaa attagtctac 900 aatgctttgg aaacagccac tggtggcttt gtcttgttgt atgagtgggt ccttcagtgg 960 cagaaaaaaa tgggtccatt ccttaccagt caagaaaaag agaagattga taagtgcaaa 1020 aagcagattc aaggaacaga aacagaattc aactcactgg taaaattgag ccatccaaat 1080 gtagtacgct accttgcaat gaatctcaaa gagcaagacg actccattgt ggtggacatt 1140 ttagtggagc acattagtgg ggtctctctt gctgcacacc tgagccactc aggccccatc 1200 cctgtgcatc agcttcgcag gtacacagct cagctcctgt caggccttga ttatctgcac 1260 agcaattctg tggtgcataa ggtcctgagt gcatctaatg tcttggtgga tgcagaaggc 1320 accgtcaaga ttacggacta tagcatttct aagcgcctcg cagacatttg caaggaggat 1380 gtgtttgagc aaacccgagt tcgttttagt gacaatgctc tgccttataa aacggggaag 1440 aaaggagatg tttggcgtct tggccttctg ctgctgtccc tcagccaagg acaggaatgt 1500 ggagagtacc ctgtgaccat ccctagtgac ttaccagctg actttcaaga ttttctaaag 1560 tgtgtgtgct tggatgacaa ggaaagatgg agtccccagc agttgttgaa acacagcttt 1620 ataaatcccc agccaaaaat gcctctagtg gaacaaagtc ctgaagattc tgaaggacaa 1680 gattatgttg agactgttat tcctagcaac cggctaccca gtgctgcctt ctttagtgag 1740 acacagagac agttttcccg atacttcatt gagtttgaag aattacaact tcttggtaaa 1800 ggagcttttg gagctgtcat caaggtgcag aacaagttgg acggctgctg ctacgcagtg 1860 aagcgcatcc ccatcaaccc ggccagccgg cagttccgca ggatcaaggg cgaagtgaca 1920 ctgctgtcac ggctgcacca tgagaacatt gtgcgctact acaacgcctg gatcgagcgg 1980 cacgagcggc cggcgggacc ggggacgccg cccccggact ccgggcccct ggccaaggat 2040 gaccgagctg cacgcgggca gccggcgagc gacacagacg gcctggacag cgtagaggcc 2100 gccgcgccgc cacccatcct cagcagctcg gtggagtgga gcacttcggg cgagcgctcg 2160 gccagtgccc gtttccccgc caccggcccg ggctccagcg atgacgagga cgacgacgag 2220 gacgagcacg gtggcgtctt ctcccagtcc ttcctgcctg cttcagattc tgaaagtgat 2280 attatctttg acaatgaaga tgagaacagt aaaagtcaga atcaggatga agattgcaat 2340 gaaaagaatg gctgccatga aagtgagcca tcagtgacga ctgaggctgt gcactaccta 2400 tacatccaga tggagtactg tgagaagagc actttacgag acaccattga ccagggactg 2460 tatcgagaca ccgtcagact ctggaggctt tttcgagaga ttctggatgg attagcttat 2520 atccatgaga aaggaatgat tcaccgggat ttgaagcctg tcaacatttt tttggattct 2580 gatgaccatg tgaaaatagg tgattttggt ttggcgacag accatctagc cttttctgct 2640 gacagcaaac aagacgatca gacaggagac ttgattaagt cagacccttc aggtcactta 2700 actgggatgg ttggcactgc tctctatgta agcccagagg tccaaggaag caccaaatct 2760 gcatacaacc agaaagtgga tctcttcagc ctgggaatta tcttctttga gatgtcctat 2820 caccccatgg tcacggcttc agaaaggatc tttgttctca accaactcag agatcccact 2880 tcgcctaagt ttccagaaga ctttgacgat ggagagcatg caaagcagaa atcagtcatc 2940 tcctggctgt tgaaccacga tccagcaaaa cggcccacag ccacagaact gctcaagagt 3000 gagctgctgc ccccacccca gatggaggag tcagagctgc atgaagtgct gcaccacacg 3060 ctgaccaacg tggatgggaa ggcctaccgc accatgatgg cccagatctt ctcgcagcgc 3120 atctcccctg ccatcgatta cacctatgac agcgacatac tgaagggcaa cttctcaatc 3180 cgtacagcca agatgcagca gcatgtgtgt gaaaccatca tccgcatctt taaaagacat 3240 ggagctgttc agttgtgtac tccactactg cttccccgaa acagacaaat atatgagcac 3300 aacgaagctg ccctattcat ggaccacagc gggatgctgg tgatgcttcc ttttgacctg 3360 cggatccctt ttgcaagata tgtggcaaga aataatatat tgaatttaaa acgatactgc 3420 atagaacgtg tgttcaggcc gcgcaagtta gatcgatttc atcccaaaga acttctggag 3480 tgtgcatttg atattgtcac ttctaccacc aacagctttc tgcccactgc tgaaattatc 3540 tacactatct atgaaatcat ccaagagttt ccagcacttc aggaaagaaa ttacagtatt 3600 tatttgaacc ataccatgtt attgaaagca atactcttac actgtgggat cccagaagat 3660 aaactcagtc aagtctacat tattctgtat gatgctgtga cagagaagct gacgaggaga 3720 gaagtggaag ctaaattttg taatctgtct ttgtcttcta atagtctgtg tcgactctac 3780 aagtttattg aacagaaggg agatttgcaa gatcttatgc caacaataaa ttcattaata 3840 aaacagaaaa caggtattgc acagttggtg aagtatggct taaaagacct agaggaggtt 3900 gttggactgt tgaagaaact cggcatcaag ttacaggtct tgatcaattt gggcttggtt 3960 tacaaggtgc agcagcacaa tggaatcatc ttccagtttg tggctttcat caaacgaagg 4020 caaagggctg tacctgaaat cctcgcagct ggaggcagat atgacctgct gattccccag 4080 tttagagggc cacaagctct ggggccagtt cccactgcca ttggggtcag catagctata 4140 gacaagatat ctgctgctgt cctcaacatg gaggaatctg ttacaataag ctcttgtgac 4200 ctcctggttg taagtgttgg ccagatgtct atgtccaggg ccatcaacct aacccagaaa 4260 ctctggacag caggcatcac agcagaaatc atgtacgact ggtcacagtc ccaagaggaa 4320 ttacaagagt actgcagaca tcatgaaatc acctatgtgg cccttgtctc ggataaagaa 4380 ggaagccatg tcaaggttaa gtctttcgag aaggaaaggc agacagagaa gcgtgtgctg 4440 gagactgaac ttgtggacca tgtactgcag aaactgagga ctaaagtcac tgatgaaagg 4500 aatggcagag aagcttccga taatcttgca gtgcaaaatc tgaaggggtc attttctaat 4560 gcttcaggtt tgtttgaaat ccatggagca acagtggttc ccattgtgag tgtgctagcc 4620 ccggagaagc tgtcagccag cactaggagg cgctatgaaa ctcaggtaca aactcgactt 4680 cagacctccc ttgccaactt acatcagaaa agcagtgaaa ttgaaattct ggctgtggat 4740 ctacccaaag aaacaatatt acagttttta tcattagagt gggatgctga tgaacaggca 4800 tttaacacaa ctgtgaagca gctgctgtca cgcctgccaa agcaaagata cctcaaatta 4860 gtctgtgatg aaatttataa catcaaagta gaaaaaaagg tgtctgtgct atttctgtac 4920 agctatagag atgactacta cagaatctta ttttaaccct aaagaactgt cgttaacctc 4980 attcaaacag acagaggctt atactggaat aatggaatgt tgtacattca tcataattta 5040 aaattaaatt ctaagaagag gctgggtgca gtggctcaca cctttaatcc cagcactttg 5100 ggaagccaag gcaggaagac tgcttgaaac caggagtttg agaccagcct gagcaacaaa 5160 gcaagacccc atctctataa aaactaaaaa aattagttgg gcatggtggc acatgcctgt 5220 agtcccagct actccagagg gtgagatgga tcatctgagc ctcaggaggt tgaggctgca 5280 gttaactgga cggcgggggg atcca

5305 34 3269 DNA Homo sapiens misc_feature Incyte ID No 1834248CB1 34 ccgggacccg gagaagatgt cttcgcggac ggtgctggcc ccgggcaacg atcggaactc 60 ggacacgcat ggcaccttgg gcagtggccg ctcctcggac aaaggcccgt cctggtccag 120 ccgctcactg ggtgcccgtt gccggaactc catcgcctcc tgtcccgagg agcagcccca 180 cgtgggcaac taccgcctgc tgaggaccat tgggaagggc aactttgcca aagtcaagct 240 ggctcggcac atcctcactg gtcgggaggt tgccatcaag attatcgaca aaacccagct 300 gaatcccagc agcctgcaga agctgttccg agaagtccgc atcatgaagg gcctaaacca 360 ccccaacatc gtgaagctct ttgaggtgat tgagactgag aagacgctgt acctggtgat 420 ggagtacgca agtgctggag aagtgtttga ctacctcgtg tcgcatggcc gcatgaagga 480 gaaggaagct cgagccaagt tccgacagat tgtttcggct gtgcactatt gtcaccagaa 540 aaatattgta cacagggacc tgaaggctga gaacctcttg ctggatgccg aggccaacat 600 caagattgct gactttggct tcagcaacga gttcacgctg ggatcgaagc tggacacgtt 660 ctgcgggagc cccccatatg ccgccccgga gctgtttcag ggcaagaagt acgacgggcc 720 ggaggtggac atctggagcc tgggagtcat cctgtacacc ctcgtcagcg gctccctgcc 780 cttcgacggg cacaacctca aggagctgcg ggagcgagta ctcagaggga agtaccgggt 840 ccctttctac atgtcaacag actgtgagag catcctgcgg agatttttgg tgctgaaccc 900 agctaaacgc tgtactctcg agcaaatcat gaaagacaaa tggatcaaca tcggctatga 960 gggtgaggag ttgaagccat acacagagcc cgaggaggac ttcggggaca ccaagagaat 1020 tgaggtgatg gtgggtatgg gctacacacg ggaagaaatc aaagagtcct tgaccagcca 1080 gaagtacaac gaagtgaccg ccacctacct cctgctgggc aggaagactg aggagggtgg 1140 ggaccggggc gccccagggc tggccctggc acgggtgcgg gcgcccagcg acaccaccaa 1200 cggaacaagt tccagcaaag gcaccagcca cagcaaaggg cagcggagtt cctcttccac 1260 ctaccaccgc cagcgcaggc atagcgattt ctgtggccca tcccctgcac ccctgcaccc 1320 caaacgcagc ccgacgagca cgggggaggc ggagctgaag gaggagcggc tgccaggccg 1380 gaaggcgagc tgcagcaccg cggggagtgg gagtcgaggg ctgcccccct ccagccccat 1440 ggtcagcagc gcccacaacc ccaacaaggc agagatccca gagcggcgga aggacagcac 1500 gagcaccccc aacaacctcc ctcctagcat gatgacccgc agaaacacct acgtttgcac 1560 agaacgcccg ggggctgagc gcccgtccct gttgccaaat gggaaagaaa acagctcagg 1620 caccccacgg gtgccccctg cctccccctc cagtcacagc ctggcacccc catcagggga 1680 gcggagccgc ctggcacgcg gttccaccat ccgcagcacc ttccatggtg gccaggtccg 1740 ggaccggcgg gcagggggtg ggggtggtgg gggtgtgcag aatgggcccc ctgcctctcc 1800 cacactggcc catgaggctg cacccctgcc cgccgggcgg ccccgcccca ccaccaacct 1860 cttcaccaag ctgacctcca aactgacccg aagggtcgca gacgaacctg agagaatcgg 1920 gggacctgag gtcacaagtt gccatctacc ttgggatcaa acggaaaccg ccccccggct 1980 gctccgattc ccctggagtg tgaagctgac cagctcgcgc cctcctgagg ccctgatggc 2040 agctctgcgc caggccacag cagccgcccg ctgccgctgc cgccagccac agccgttcct 2100 gctggcctgc ctgcacgggg gtgcgggcgg gcccgagccc ctgtcccact tcgaagtgga 2160 ggtctgccag ctgccccggc caggcttgcg gggagttctc ttccgccgtg tggcgggcac 2220 cgccctggcc ttccgcaccc tcgtcacccg catctccaac gacctcgagc tctgagccac 2280 cacggtccca gggcccttac tcttcctctc ccttgtcgcc ttcacttcta caggagggga 2340 aggggccagg gaggggattc tccctttatc atcacctcag tttccctgaa ttatatttgg 2400 gggcaaagat tgtcccctct gctgttctct ggggccgctc agcacagaag aaggatgagg 2460 gggctcagcg gggggagctg gcaccttcct ggagcctcca gccagtcctg tcctccctcg 2520 ccctaccaag agggcacctg aggagacttt ggggacaggg caggggcagg gagggaaact 2580 gaggaaatct tccattcctc ccaacagctc aaaattaggc cttgggcagg ggcagggaga 2640 gctgctgagc ctaaagactg gagaatctgg gggactggga gtgggggtca gagaggcaga 2700 ttccttcccc tcccgtcccc tcacgctcaa acccccactt cctgccccag gctggcgcgg 2760 ggcactttgt acaaatcctt gtaaataccc cacaccctcc cctctgcaaa ggtctcttga 2820 ggagctgccg ctgtcaccta cggtttttaa gttattacac cccgaccctc ctcctgtcag 2880 ccccctcacc tgcagcctgt tgcccaataa atttaggaga gtccccccct ccccaatgct 2940 gaccctagga ttttccttcc ctgccctcac ctgcaaatga gttaaagaag aggcgtggga 3000 atccaggcag tggtttttcc tttcggagcc tcggttttct catctgcaga atgggagcgg 3060 tgggggtggg aaggtaagga tggtcgtgga agaaggcagg atggaactcg gcctcatccc 3120 cgaggcccca gttcctatat cgggcccccc attcatccac tcacactccc agccaccatg 3180 ttacactgga ctctaagcca cttcttactc cagtagtaaa tttattcaat aaacaatcat 3240 tgacccatga aaaaaaaaaa aaaaaaaaa 3269 35 3017 DNA Homo sapiens misc_feature Incyte ID No 71584520CB1 35 gggcaagcgg ctggcgatgc tggaggttcg ctagccgaag cggctgcatc tggcgccgcg 60 tctgccccgc gtgctcggag cggattctgc ccgccgtccc cggagccctc ggcgccccgc 120 tgagcccggc gatcacttcc tccctgtgac caaccggcgc tgcaggttag agcctggcaa 180 tgccgtttgg gtgtgtgact ctgggcgaca agaagaacta taaccagcca tcggaggtga 240 ctgacagata tgatttggga caggtcatca agactgagga gttttgtgaa atcttccggg 300 ccaaggacaa gacgacaggc aagctgcaca cctgcaagaa gttccagaag cgggacggcc 360 gcaaggtgcg gaaagctgcc aagaacgaga taggcatcct caagatggtg aagcatccca 420 acatcctaca gctggtggat gtgtttgtga cccgcaagga gtactttatc ttcctggagc 480 tggccacggg gagggaggtg tttgactgga tcctggacca gggctactac tcggagcgag 540 acacaagcaa cgtggtacgg caagtcctgg aggccgtggc ctatttgcac tcactcaaga 600 tcgtgcacag gaatctcaag ctggagaacc tggtttacta caaccggctg aagaactcga 660 agattgtcat cagtgacttc catctggcta agctagaaaa tggcctcatc aaggagccct 720 gtgggacccc cgagtatctg gccccagagg tggtaggccg gcagcggtat ggacgccctg 780 tggactgctg ggccattgga gtcatcatgt acatcctgct ttcaggcaac ccacctttct 840 atgaggaggt ggaagaagat gattatgaga accatgataa gaatctcttc cgcaagatcc 900 tggctggtga ctatgagttt gactctccat attgggatga tatttcgcag gcagccaaag 960 acctggtcac aaggctgatg gaggtggagc aagaccagcg gatcactgca gaagaggcca 1020 tctcccatga gtggatttct ggcaatgctg cttctgataa gaacatcaag gatggtgtct 1080 gtgcccagat tgaaaagaac tttgccaggg ccaagtggaa gaaggctgtc cgagtgacca 1140 ccctcatgaa acggctccgg gcaccagagc agtccagcac ggctgcagcc cagtcggcct 1200 cagccacaga cactgccacc cccggggctg caggtggggc cacagctgca gctgcgagtg 1260 gagctacctc agcccctgag ggtgatgctg ctcgtgctgc aaagagtgat aatgtggccc 1320 ccgcagaccg tagtgccacc ccagccacag atggaagtgc caccccagcc actgatggca 1380 gtgtcacccc agccaccgat ggaagcatca ctccagccac tgatgggagt gtcaccccag 1440 ccactgacag gagcgctact ccagccactg atgggagagc cacaccagcc acagaagaga 1500 gcactgtgcc caccacccaa agcagtgcca tgctggccac caaggcagct gccacccctg 1560 agccggctat ggcccagccg gacagcacag ccccagaggg cgccacaggc caggctccac 1620 cctctagtaa aggggaagag gctgctggtt atgcccagga gtctcaaagg gaggaggcca 1680 gctgagtagg cagcctggtg agggggggca ggggatgggc aggagggtgg gagagtggat 1740 gaggggcttc tcactgtaca tagagtcact ggcatgatgc cctcgctccc ccatgccccc 1800 acatcccagt ggggcataac taggggtcac gggagagcag tctcgtctcc tgtgtgtatg 1860 tgtgtgagtg gtgggcaggc cagtggcagg gccggcccca gcccctgcat ggattccttg 1920 tggcttttct gtcttttgct agcttcacca gtttctgttc cttgtgggat gctgctctag 1980 ggatactcag ggggctcctg ctctccttcc ccttcccttc ttgcctcacc attcccctag 2040 gcaggccctg caggtcccac actctcccag gccctaaact tgggcggcct tgccctgaga 2100 gctggtcctc cagcgaggcc ctgtcagcgg tcttaggctc ctgcacatga aggtgtgtgc 2160 ctgtggtgtg tgggctgctc taggagcaga tacaggctgg tatagaggat gcagaaaggt 2220 agggcagtat gtttaagtcc agacttggca catggctagg gatactgctc actagctgtg 2280 gaggtcctca ggagtggaga gaatgagtag gagggcagaa gcttccattt ttgtccttcc 2340 taagaccctg ttatttgtgt tatttcctgc ctttccgagt cctgcagtgg gctgccctgt 2400 accctgaacc tcatgagcct ctaagggaaa ggaggaacaa ttaggacgtg gcaatgagac 2460 ctggcagggc agagtacaag cccagcaccc agtgtcccag ccttactggg tccttaccct 2520 gggccaaaca gggagggctg atacctcctt gctcttccta gatgcccacc tcctacaatc 2580 tcagcccaca agtcctctcc accctagggg gcttgctgca tggcaataac tcataatctg 2640 atttggaggt ttgcccttta caggggcaga ttttctgctc agttcaacaa tgaaatgaag 2700 aggaactccc tctttctaca gctcacttct atcagaggcc caggtgcctc agagccacat 2760 tgagttgctt tttctgggat gaggaagtag ggttaaactc cccagtttcc tgagggaggc 2820 tcctgacagg tgccctttgt cagaccctac cacagcctgg ataggcagcc acattggtcc 2880 tcgcccttgc tcggcactcc gtggtggtcc tgcccttctc cctgcatgcc tgtgggtctg 2940 ctctggtgtg tgaaggtcgg tgggttaact gtgtgcctac tgaacctggc aaataaacat 3000 caccctgcaa agccaaa 3017 36 2168 DNA Homo sapiens misc_feature Incyte ID No 7475538CB1 36 gctgtcatcg ttccgtgggc cctgcttgcg ggcacgctct cggcgcatgc gttttttatg 60 cgggattaag cttgctgctg cgtgacagcg gagggctagg aaaaggcgca gtggggcccg 120 gagctgtcac ccctgactcg acgcagcttc cgttctcctg gtgacgtggc ctacaggaac 180 cgccccagtg gtcagctgcc gcgctgttgc taggcaacag cgtgcgagct cagatcagcg 240 tggggtggag gagaagtgga gtttggaagt tcaggggcac aggggcacag gcccacgact 300 gcagcgggat ggaccagtac tgcatcctgg gccgcatcgg ggagggcgcc cacggaatcg 360 tcttcaaggc caagcacgtg gagactggcg agatagttgc cctcaagaag gtggccctaa 420 ggcggttgga agacggcttc cctaaccagg ccctgcggga gattaaggct ctgcaggaga 480 tggaggacaa tcagtatgtg gtacaactga aggctgtgtt cccacacggt ggaggctttg 540 tgctggcctt tgagttcatg ctgtcggatc tggccgaggt ggtgcgccat gcccagaggc 600 cactagccca ggcacaggtc aagagctacc tgcagatgct gctcaagggt gtcgccttct 660 gccatgccaa caacattgta catcgggacc tgaaacctgc caacctgctc atcagcgcct 720 caggccagct caagatagcg gactttggcc tggctcgagt cttttcccca gacggcagcc 780 gcctctacac acaccaggtg gccaccaggt ggtaccgagc ccccgagctc ctgtatggtg 840 cccgccagta tgaccagggc gtcgatctgt ggtctgtggg ctgcatcatg ggggagctgt 900 tgaatgggtc cccccttttc ccgggcaaga acgatattga acagctttgc tatgtgcttc 960 gcatcttggg caccccaaac cctcaagtct ggccggagct cactgagctg ccggactaca 1020 acaagatctc ctttaaggag caggtgccca tgcccctgga ggaggtgctg cctgacgtct 1080 ctccccaggc attggatctg ctgggtcaat tccttctcta ccctcctcac cagcgcatcg 1140 cagcttccaa ggctctcctc catcagtact tcttcacagc tcccctgcct gcccatccat 1200 ctgagctgcc gattcctcag cgtctagggg gacctgcccc caaggcccat ccagggcccc 1260 cccacatcca tgacttccac gtggaccggc ctcttgagga gtcgctgttg aacccagagc 1320 tgattcggcc cttcatcctg gaggggtgag aagttggccc tggtcccgtc tgcctgctcc 1380 tcaggaccac tcagtccacc tgttcctctg ccacctgcct ggcttcaccc tccaaggcct 1440 ccccatggcc acagtgggcc cacaccacac cctgcccctt agcccttgcg agggttggtc 1500 tcgaggcaga ggtcatgttc ccagccaaga gtatgagaac atccagtcga gcagaggaga 1560 ttcatggcct gtgctcggtg agccttacct tctgtgtgct actgacgtac ccatcaggac 1620 agtgagctct gctgccagtc aaggcctgca tatgcagaat gacgatgcct gccttggtgc 1680 tgcttccccg agtgctgcct cctggtcaag gagaagtgca gagagtaagg tgtccttatg 1740 ttggaaactc aagtggaagg aagatttggt ttggttttat tctcagagcc attaaacact 1800 agttcagtat gtgagatata gattctaaaa acctcaggtg gctctgcctt atgtctgttc 1860 ctccttcatt tctctcaagg gaaatggcta aggtggcatt gtctcatggc tctcgttttt 1920 ggggtcatgg ggagggtagc accaggcata gccacttttg ccctgaggga ctcctgtgtg 1980 cttcacatca ctgagcactc atttagaagt gagggagaca gaagtctagg cccagggatg 2040 gctccagttg gggatccagc aggagaccct ctgcacatga ggctggttta ccaacatcta 2100 ctccctcagg atgagcgtga gccagaagca gctgtgtatt taaggaaaca agcgttcctg 2160 gaattaat 2168

Claims

What is claimed is:

1. An isolated polypeptide selected from the group consisting of: a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQID NO:1-18, b) a naturally occurring polypeptide comprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:2-18, c) a naturally occurring polypeptide comprising an amino acid sequence at least 98% identical to an amino acid sequence of SEQ ID NO:1, d) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, and e) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18.

2. An isolated polypeptide of claim 1 selected from the group consisting of SEQ ID NO:1-18.

3. An isolated polynucleotide encoding a polypeptide of claim 1.

4. An isolated polynucleotide encoding a polypeptide of claim 2.

5. An isolated polynucleotide of claim 4 selected from the group consisting of SEQ ID NO:19-36.

6. A recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide of claim 3.

7. A cell transformed with a recombinant polynucleotide of claim 6.

8. A transgenic organism comprising a recombinant polynucleotide of claim 6.

9. A method for producing a polypeptide of claim 1, the method comprising: a) culturing a cell under conditions suitable for expression of the polypeptide, wherein said cell is transformed with a recombinant polynucleotide, and said recombinant polynucleotide comprises a promoter sequence operably linked to a polynucleotide encoding the polypeptide of claim 1, and b) recovering the polypeptide so expressed.

10. An isolated antibody which specifically binds to a polypeptide of claim 1.

11. An isolated polynucleotide selected from the group consisting of: a) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:19-36, b) a naturally occurring polynucleotide comprising a polynucleotide sequence at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NO:20-36, c) a naturally occurring polynucleotide comprising a polynucleotide sequence at least 98% identical to the polynucleotide sequence of SEQ ID NO:19, d) a polynucleotide complementary to a polynucleotide of a), e) a polynucleotide complementary to a polynucleotide of b), f) a polynucleotide complementary to a polynucleotide of c), and g) an RNA equivalent of a)-f).

12. An isolated polynucleotide comprising at least 60 contiguous nucleotides of a polynucleotide of claim 11.

13. A method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide of claim 11, the method comprising: a) hybridizing the sample with a probe comprising at least 20 contiguous nucleotides comprising a sequence complementary to said target polynucleotide in the sample, and which probe specifically hybridizes to said target polynucleotide, under conditions whereby a hybridization complex is formed between said probe and said target polynucleotide or fragments thereof, and b) detecting the presence or absence of said hybridization complex, and, optionally, if present, the amount thereof.

14. A method of claim 13, wherein the probe comprises at least 60 contiguous nucleotides.

15. A method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide of claim 11, the method comprising: a) amplifying said target polynucleotide or fragment thereof using polymerase chain reaction amplification, and b) detecting the presence or absence of said amplified target polynucleotide or fragment thereof, and, optionally, if present, the amount thereof.

16. A composition comprising a polypeptide of claim 1 and a pharmaceutically acceptable excipient.

17. A composition of claim 16, wherein the polypeptide has an amino acid sequence selected from the group consisting of SEQ ID NO:1-18.

18. A method for treating a disease or condition associated with decreased expression of functional PKIN, comprising administering to a patient in need of such treatment the composition of claim 16.

19. A method for screening a compound for effectiveness as an agonist of a polypeptide of claim 1, the method comprising: a) exposing a sample comprising a polypeptide of claim 1 to a compound, and b) detecting agonist activity in the sample.

20. A composition comprising an agonist compound identified by a method of claim 19 and a pharmaceutically acceptable excipient.

21. A method for treating a disease or condition associated with decreased expression of functional PKIN, comprising administering to a patient in need of such treatment a composition of claim 20.

22. A method for screening a compound for effectiveness as an antagonist of a polypeptide of claim 1, the method comprising: a) exposing a sample comprising a polypeptide of claim 1 to a compound, and b) detecting antagonist activity in the sample.

23. A composition comprising an antagonist compound identified by a method of claim 22 and a pharmaceutically acceptable excipient.

24. A method for treating a disease or condition associated with overexpression of functional PKIN, comprising administering to a patient in need of such treatment a composition of claim 23.

25. A method of screening for a compound that specifically binds to the polypeptide of claim 1, said method comprising the steps of: a) combining the polypeptide of claim 1 with at least one test compound under suitable conditions, and b) detecting binding of the polypeptide of claim 1 to the test compound, thereby identifying a compound that specifically binds to the polypeptide of claim 1.

26. A method of screening for a compound that modulates the activity of the polypeptide of claim 1, said method comprising: a) combining the polypeptide of claim 1 with at least one test compound under conditions permissive for the activity of the polypeptide of claim 1, b) assessing the activity of the polypeptide of claim 1 in the presence of the test compound, and c) comparing the activity of the polypeptide of claim 1 in the presence of the test compound with the activity of the polypeptide of claim 1 in the absence of the test compound, wherein a change in the activity of the polypeptide of claim 1 in the presence of the test compound is indicative of a compound that modulates the activity of the polypeptide of claim 1.

27. A method for screening a compound for effectiveness in altering expression of a target polynucleotide, wherein said target polynucleotide comprises a sequence of claim 5, the method comprising: a) exposing a sample comprising the target polynucleotide to a compound, under conditions suitable for the expression of the target polynucleotide, b) detecting altered expression of the target polynucleotide, and c) comparing the expression of the target polynucleotide in the presence of varying amounts of the compound and in the absence of the compound.

28. A method for assessing toxicity of a test compound, said method comprising: a) treating a biological sample containing nucleic acids with the test compound; b) hybridizing the nucleic acids of the treated biological sample with a probe comprising at least 20 contiguous nucleotides of a polynucleotide of claim 11 under conditions whereby a specific hybridization complex is formed between said probe and a target polynucleotide in the biological sample, said target polynucleotide comprising a polynucleotide sequence of a polynucleotide of claim 11 or fragment thereof; c) quantifying the amount of hybridization complex; and d) comparing the amount of hybridization complex in the treated biological sample with the amount of hybridization complex in an untreated biological sample, wherein a difference in the amount of hybridization complex in the treated biological sample is indicative of toxicity of the test compound.

29. A diagnostic test for a condition or disease associated with the expression of PKIN in a biological sample comprising the steps of: a) combining the biological sample with an antibody of claim 10, under conditions suitable for the antibody to bind the polypeptide and form an antibody:polypeptide complex; and b) detecting the complex, wherein the presence of the complex correlates with the presence of the polypeptide in the biological sample.

30. The antibody of claim 10, wherein the antibody is: a) a chimeric antibody, b) a single chain antibody, c) a Fab fragment, d) a F(ab').sub.2 fragment, or e) a humanized antibody.

31. A composition comprising an antibody of claim 10 and an acceptable excipient.

32. A method of diagnosing a condition or disease associated with the expression of PKN in a subject, comprising administering to said subject an effective amount of the composition of claim 31.

33. A composition of claim 31, wherein the antibody is labeled.

34. A method of diagnosing a condition or disease associated with the expression of PKIN in a subject, comprising administering to said subject an effective amount of the composition of claim 33.

35. A method of preparing a polyclonal antibody with the specificity of the antibody of claim 10 comprising: a) immunizing an animal with a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, or an immunogenic fragment thereof, under conditions to elicit an antibody response; b) isolating antibodies from said animal; and c) screening the isolated antibodies with the polypeptide, thereby identifying a polyclonal antibody which binds specifically to a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18.

36. An antibody produced by a method of claim 35.

37. A composition comprising the antibody of claim 36 and a suitable carrier.

38. A method of making a monoclonal antibody with the specificity of the antibody of claim comprising: a) immunizing an animal with a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18, or an immunogenic fragment thereof, under conditions to elicit an antibody response; b) isolating antibody producing cells from the animal; c) fusing the antibody producing cells with immortalized cells to form monoclonal antibody-producing hybridoma cells; d) culturing the hybridoma cells; and e) isolating from the culture monoclonal antibody which binds specifically to a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18.

39. A monoclonal antibody produced by a method of claim 38.

40. A composition comprising the antibody of claim 39 and a suitable carrier.

41. The antibody of claim 10, wherein the antibody is produced by screening a Fab expression library.

42. The antibody of claim 10, wherein the antibody is produced by screening a recombinant immunoglobulin library.

43. A method for detecting a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18 in a sample, comprising the steps of: a) incubating the antibody of claim 10 with a sample under conditions to allow specific binding of the antibody and the polypeptide; and b) detecting specific binding, wherein specific binding indicates the presence of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18 in the sample.

44. A method of purifying a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18 from a sample, the method comprising: a) incubating the antibody of claim 10 with a sample under conditions to allow specific binding of the antibody and the polypeptide; and b) separating the antibody from the sample and obtaining the purified polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:1-18.

45. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:1.

46. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:2.

47. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:3.

48. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:4.

49. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:5.

50. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:6.

51. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:7.

52. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:8.

53. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:9.

54. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:10.

55. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:11.

56. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:12.

57. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:13.

58. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:14.

59. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:15.

60. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:16.

61. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:17.

62. A polypeptide of claim 1, comprising the amino acid sequence of SEQ ID NO:18.

63. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:19.

64. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:20.

65. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:21.

66. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:22.

67. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:23.

68. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:24.

69. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:25.

70. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:26.

71. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:27.

72. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:28.

73. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:29.

74. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:30.

75. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:31.

76. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:32.

77. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:33.

78. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:34.

79. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:35.

80. A polynucleotide of claim 11, comprising the polynucleotide sequence of SEQ ID NO:36.